Safe and stable nimodipine for injection and a preparation method thereof

Abstract

The present application relates to the technical field of medicine, in particular to a safe and stable nimodipine for injection and a preparation method thereof, and provides a nimodipine cyclodextrin freeze-dried powder injection containing sulfobutyl ether-beta-cyclodextrin, which can significantly improve the solubility, drug safety and dilution stability of nimodipine, increase the compliance of patients and the convenience of clinical use. Compared with the commercially available nimodipine injection, the safety and dilution stability of the present application are significantly improved, and the present application has obvious advantages.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of medicine, in particular, a safe and stable nimodipine for injection and a preparation method thereof. BACKGROUND

[0002] Nimodipine (structural formula as shown in formula I) is a yellow crystalline powder with a melting point of 125℃, which is almost insoluble in water, soluble in ethanol and chloroform, and easily degradable under light. Nimodipine can easily penetrate the blood-brain barrier, act on cerebral vascular smooth muscle, dilate cerebral blood vessels, and increase cerebral blood flow, which can effectively prevent or reverse ischemic damage of brain tissue caused by cerebral vasospasm after subarachnoid hemorrhage. It is clinically used for the prevention and treatment of ischemic cerebrovascular diseases, such as ischemic nerve damage caused by cerebral vasospasm after subarachnoid hemorrhage, migraine, sudden deafness, etc., and has great potential in the treatment of cerebrovascular diseases (Carlson AP, et al. Nimodipine Reappraised: An Old Drug With a Future. Curr Neuropharmacol. 2020; 18(1): 65-82).

[0003]

[0004] At present, the commercially available dosage forms of nimodipine include tablets, capsules, oral solutions and injections. After oral administration of nimodipine, the liver first-pass effect is significant, and the bioavailability is only 5-15%; and the half-life is short, so patients need to take frequent medication to maintain effective blood drug concentration, which greatly limits the oral administration. (Teng Z, et al. Preparation and characterization of nimodipine-loaded nanostructured lipid systems for enhanced solubility and bioavailability. Int J Nanomedicine. 2018; 14: 119-133.)

[0005] In order to improve the defects of oral nimodipine preparation, Bayer Company of Germany developed a nimodipine injection. Nimodipine injection has no liver first-pass effect, high bioavailability, rapid onset, and other advantages that other dosage forms cannot match. However, the injection has the following problems: ① Because of the poor water solubility of nimodipine, a large amount of organic solvent is added to the nimodipine injection to increase the solubility of nimodipine, including 23.7% (V / V) ethanol and 17% (V / V) polyethylene glycol 400. A large amount of organic solvent has a strong irritation to blood vessels, causing pain, redness, and other symptoms at the injection site, and easily causing phlebitis, resulting in poor patient compliance (Schneider AC, et al. Chronic exposure to ethanol causes steatosis and inflammation in zebrafish liver. World J Hepatol. 2017; 9(8): 418-426.). ② Nimodipine injection is prone to precipitation when diluted directly for clinical use. Therefore, when administered, a three-way valve is used to mix with 5% glucose, 0.9% sodium chloride, and other liquids at a ratio of 1:4 (V / V) for infusion, with a daily administration time of 10 h, a treatment time of 10-14 days. This infusion method has a high safety risk, and the infusion speed and mixing ratio are not easy to control, the operation is cumbersome and time-consuming, and the clinical use is very inconvenient, with poor patient compliance.

[0006] In view of the shortcomings of existing nimodipine preparations, researchers in the industry have carried out extensive research on nimodipine.

[0007] In order to improve the solubility of nimodipine, researchers have used nanometer preparation technology such as fat emulsion, liposome, and lipid nanoparticle to encapsulate nimodipine, such as Chinese patent documents CN107019682A, CN102552156A, CN101199522A, CN1554340A, and CN105434355A. Although the water solubility of nimodipine is improved, the nanometer preparation has the problems of easy aggregation of microparticles, low drug loading, low encapsulation efficiency, and obvious burst effect, and is easily captured by the liver and spleen reticuloendothelial system, affecting the efficacy and even increasing the toxicity. In addition, the preparation process of nanometer preparation is complex, the production line requires high, the cost is high, and the product quality is difficult to control.

[0008] There are also researchers using surfactants, polymers and other with nimodipine micelles, thereby increasing its water solubility. Such as Chinese patent document CN101129366A, CN1771950A, its formula using Tween 80 and other highly toxic surfactants, easy to cause hemolysis and allergic reactions, strong irritability; Chinese patent document CN113694031A, CN102525917A, CN103315948A using polymer coated nimodipine, polymer micelles in vivo process is difficult to predict, affect the efficacy.

[0009] In view of the above deficiencies of nano-preparation, researchers began to look for new preparation technology to improve the water solubility of nimodipine, among which nimodipine cyclodextrin inclusion complex is one of the current research hotspots, and domestic and foreign scholars have carried out a lot of research.

[0010] Literature reports that nimodipine is difficult to form inclusion complex with β-cyclodextrin due to the large steric hindrance of the side chain (Wu Xuefen et al. Interaction between β-cyclodextrin and dihydropyridine drugs [J]. Chinese Journal of Pharmacy, 2005 (08): 599-601.). In view of the shortcomings of β-cyclodextrin, researchers try to find new β-cyclodextrin derivatives to load nimodipine.

[0011] Chinese patent document CN1634050A discloses a new nimodipine composition for injection, which is composed of nimodipine, polyethylene glycol 400, Tween 80 and hydroxypropyl-β-cyclodextrin. This technology has some obvious defects: ①The formula contains hydroxypropyl-β-cyclodextrin, and a large amount of Tween 80 and polyethylene glycol 400 are added to help dissolution, and the prescription process is complex. ②The addition of polyethylene glycol 400 in the freeze-dried product leads to the appearance of shrinkage, collapse and unsatiation; ③The formula contains Tween 80, which is easy to cause hemolysis and allergic reactions, and has strong irritability.

[0012] Chinese patent document CN1424035A discloses a nimodipine freeze-dried composition, which contains phospholipids, cyclodextrin and its derivatives or surfactants, wherein the mass ratio of nimodipine to cyclodextrin and its derivatives is 1:1 to 1:20. Chinese patent document CN1653089A discloses a complex of organic drugs and β-cyclodextrin derivatives and its preparation method, wherein the mass ratio of nimodipine to hydroxypropyl-β-cyclodextrin is 1:136.

[0013] The above patents generally have the following problems: ① The relative proportion of hydroxypropyl-β-cyclodextrin and drug is too high, and the inclusion effect is poor. ② A large amount of organic solvent is added in the formula to assist dissolution, which is highly irritating. ③ The presence of a large amount of organic solvent is not conducive to cyclodextrin inclusion. In summary, solving the technical defects of the above cyclodextrin inclusion compounds is still a hot research topic. In recent years, sulfobutyl ether-β-cyclodextrin-containing injections have been successively listed, and the application of sulfobutyl ether-β-cyclodextrin in the field of pharmaceutical preparations has attracted widespread attention from researchers. Literature reports that the solubilization effect of sulfobutyl ether-β-cyclodextrin on nimodipine is significantly better than that of hydroxypropyl-β-cyclodextrin, and compared with hydroxypropyl-β-cyclodextrin, sulfobutyl ether-β-cyclodextrin is a more ideal carrier for nimodipine. (Semcheddine F, et al. Effects of the Preparation Method on the Formation of True Nimodipine SBE-β-CD / HP-β-CD Inclusion Complexes and Their Dissolution Rates Enhancement. AAPS PharmSciTech. 2015; 16(3): 704-715.)

[0014] Literature search found that some scholars used grinding method to prepare nimodipine sulfobutyl ether-β-cyclodextrin inclusion complex, the drug loading of the inclusion complex prepared by this method was low, and after preparation, it still needed to be filtered and washed to remove the undissolved drug, which had poor repeatability, the product quality was difficult to guarantee, and was not suitable for industrial production(Yang X, Ren Y. Study on the properties of nimodipine / sulfobutyl ether-β-cyclodextrin solid inclusion complex [J]. China modern applied pharmacy, 2003(05): 380-383.). Many researchers used shaking method to prepare nimodipine sulfobutyl ether-β-cyclodextrin inclusion complex. Serena et al. mixed excess nimodipine into 50 mM sulfobutyl ether-β-cyclodextrin aqueous solution, and shook on a shaking table at room temperature for 48 h to prepare nimodipine sulfobutyl ether-β-cyclodextrin inclusion complex(Tongiani S, et al. Sulfobutyl ether-alkyl ether mixed cyclodextrin derivatives with enhanced inclusion ability. J Pharm Sci. 2009; 98(12): 4769-4780.); Farouk et al. mixed excess nimodipine into 16 mM sulfobutyl ether-β-cyclodextrin aqueous solution, and shook on a shaking table at 37℃ for 72 h to prepare nimodipine sulfobutyl ether-β-cyclodextrin inclusion complex(Semcheddine F, et al. Effects of the Preparation Method on the Formation of True Nimodipine SBE-β-CD / HP-β-CD Inclusion Complexes and Their Dissolution Rates Enhancement. AAPS PharmSciTech. 2015; 16(3): 704-715.); Doaa Nabih et al. mixed excess nimodipine into sulfobutyl ether-β-cyclodextrin aqueous solution, vortexed the suspension for 5 min, then ultrasonicated for 15 min, and finally shook on a shaking table at 37℃ for 72 h to prepare nimodipine sulfobutyl ether-β-cyclodextrin inclusion complex(Maria DN, et al. Nimodipine ophthalmic formulations for management of glaucoma. Pharm Res. 2017; 34(4): 809-824.).The above documents all use a shaking method to prepare the inclusion compound, and the inclusion effect of the cyclodextrin inclusion compound prepared by the method is poor. In addition, since these documents do not systematically study the amount of sulfobutyl ether-β-cyclodextrin, the inclusion temperature, the inclusion time, the amount of ethanol, the mass ratio of sulfobutyl ether-β-cyclodextrin and nimodipine (the ratio of adjuvant), and the stirring speed, the problems of long preparation time, low solubility of nimodipine, and high adjuvant ratio are generally present.

[0015] In summary, the prior art generally has poor cyclodextrin inclusion effect, long preparation time, and high adjuvant ratio. In addition to containing cyclodextrin, a large amount of organic solvent is added to assist dissolution in the formula, which is not conducive to cyclodextrin inclusion, increases the irritability and toxicity of the preparation, and has low safety. Therefore, it is of great significance to develop a safe and stable nimodipine injection, which can lay a solid foundation for improving the safety and convenience of nimodipine clinical medication. SUMMARY

[0016] Based on the above prior art, the defects of poor dilution stability and low safety of nimodipine injection have not been fundamentally changed. The purpose of the present application is to provide a safe and stable nimodipine injection and a preparation method thereof.

[0017] The present application increases the solubility of nimodipine by using sulfobutyl ether-β-cyclodextrin. Based on the preliminary literature research and experimental verification, it is found that sulfobutyl ether-β-cyclodextrin is an ideal carrier for nimodipine. However, there are few studies on nimodipine sulfobutyl ether-β-cyclodextrin inclusion compounds, and a nimodipine sulfobutyl ether-β-cyclodextrin inclusion compound product suitable for industrial production and applicable to clinical use has not been developed.

[0018] The inventors prepared a nimodipine lyophilized powder according to Chinese patent document CN1634050A. The formula contains Tween 80, which easily leads to hemolysis and allergic reactions, and has strong irritability. In addition, the lyophilized powder obtained has loose voids and collapses (see Example 1), and the appearance of unqualified lyophilized powder affects the quality of the product.

[0019] The inventors prepared a nimodipine powder containing 2-hydroxypropyl-β-cyclodextrin according to Example 2 of Chinese patent document CN1424035A. It was found through multiple experiments that the drug precipitated during preparation. Even if the nimodipine cyclodextrin inclusion compound was prepared according to the largest amount of adjuvant in the patent (drug / 2-hydroxypropyl-β-cyclodextrin mass ratio 1:20), the drug could not be completely included, and the actual adjuvant ratio was 571 (see Example 2).

[0020] The inventors followed the operation of Example 28 in Chinese patent document CN1653089A and verified through multiple experiments that the drug precipitated during preparation. The actual adjuvant ratio was 1904 (see Example 3), which is much higher than the adjuvant ratio of the present application.

[0021] The inventors also prepared nimodipine sulfobutyl ether-β-cyclodextrin inclusion complex by the shaking method used in the literature, and the nimodipine sulfobutyl ether-β-cyclodextrin inclusion complex prepared by shaking at 37℃ for 72h had a poor inclusion effect and a long preparation time, which was not conducive to the industrial production of the injection (see Example 4).

[0022] The inventors found through a large number of experimental studies that the amount of sulfobutyl ether-β-cyclodextrin, the inclusion temperature, the inclusion time, the amount of ethanol, the auxiliary drug ratio and the stirring speed significantly affect the inclusion effect of sulfobutyl ether-β-cyclodextrin on nimodipine. The inventors can significantly reduce the auxiliary drug ratio by adjusting the amount of sulfobutyl ether-β-cyclodextrin, the inclusion temperature, the inclusion time, the amount of ethanol and the stirring speed, achieve an ideal drug loading concentration, and the safety and dilution stability of the preparation are both good. In addition, a small amount of ethanol can be contained in the formulation of the present application, and dissolving nimodipine in a small amount of ethanol can increase the solubility of the drug in sulfobutyl ether-β-cyclodextrin in aqueous solution, increase the convenience of the preparation process, and ethanol can be removed during the preparation process, leaving less residue in the product (see Examples 33 and 65).

[0023] In a first aspect of the present application, a safe and stable nimodipine injection is provided, which is prepared from sulfobutyl ether-β-cyclodextrin and nimodipine, and the mass ratio of the sulfobutyl ether-β-cyclodextrin and the nimodipine is 200:1-700:1.

[0024] Further, the mass ratio of the sulfobutyl ether-β-cyclodextrin and the nimodipine is 350:1-700:1.

[0025] Further, the nimodipine content of the nimodipine injection before lyophilization is 0.01-0.14% g / mL, and the sulfobutyl ether-β-cyclodextrin content is 10-50% g / mL.

[0026] Further, the mass ratio of the sulfobutyl ether-β-cyclodextrin and the nimodipine is 400:1-600:1, the nimodipine content is 0.02-0.10% g / mL, and the sulfobutyl ether-β-cyclodextrin concentration is 10-40% g / mL.

[0027] Further preferably, the mass ratio of the sulfobutyl ether-β-cyclodextrin and the nimodipine is 450:1-550:1, the nimodipine content is 0.04-0.06% g / mL, and the sulfobutyl ether-β-cyclodextrin concentration is 20-30% g / mL.

[0028] A safe and stable nimodipine injection of the present application is prepared from the following ingredients:

[0029]

[0030]

[0031] Further, the nimodipine injection is made of the following ingredients:

[0032]

[0033]

[0034] Further, the nimodipine injection is made of the following ingredients:

[0035]

[0036] Further, the nimodipine injection above does not contain ethanol.

[0037] Further, the nimodipine injection above can also contain a buffer to adjust the pH value to 4.5-7.5; the buffer is selected from one of the following: sodium citrate-citric acid, disodium hydrogen phosphate-citric acid, disodium hydrogen phosphate-disodium dihydrogen phosphate, disodium hydrogen phosphate-potassium dihydrogen phosphate and potassium dihydrogen phosphate-sodium hydroxide.

[0038] In the second aspect of the present application, a preparation method of the safe and stable nimodipine injection is provided, comprising the following steps:

[0039] (a) weigh the prescribed amount of sulfobutyl ether-β-cyclodextrin (and buffer), add an appropriate amount of water for injection, stir to dissolve at a certain temperature, and obtain a sulfobutyl ether-β-cyclodextrin aqueous solution;

[0040] (b) weigh the prescribed amount of nimodipine, add the prescribed amount of ethanol to dissolve, and obtain a nimodipine ethanol solution;

[0041] (c) under the conditions of a certain stirring temperature and speed, add the solution of step (b) or nimodipine drug powder to the solution of step (a), stir for a certain time, add water for injection to the full amount, sterilize with a 0.22 μm microporous filter, fill, freeze-dry, and package, and obtain the safe and stable nimodipine injection.

[0042] Further, the stirring temperature in steps (a) and (c) is 20-100℃, further preferably 40-95℃, and more preferably 60-90℃.

[0043] Further, the stirring time in step (c) is 10-300 min, further preferably 30-240 min, and more preferably 60-180 min.

[0044] Further, the stirring speed in step (c) is 0.5-10.0 m / s, further preferably 0.8-6.0 m / s, and more preferably 1.0-3.0 m / s.

[0045] Further, clean and dry nitrogen (N2) can be introduced to saturation in steps (a), (b) and (c), including nitrogen filling after freeze drying and packaging.

[0046] Further, the ethanol residue in the nimodipine for injection in step (c) is less than 0.3%, further preferably less than 0.1%, and more preferably less than 0.05%.

[0047] The present application is based on a large number of experimental studies, and it is found that the amount of sulfobutyl ether-β-cyclodextrin, the inclusion temperature, the inclusion time, the amount of ethanol, the ratio of auxiliary drugs and the stirring speed significantly affect the inclusion effect of sulfobutyl ether-β-cyclodextrin on nimodipine. By controlling the conditions of the present application, the relative proportion of sulfobutyl ether-β-cyclodextrin and nimodipine can be significantly reduced, the safety and dilution stability of the preparation can be improved, and the defects of commercially available nimodipine injection can be overcome. Therefore, the control of the amount of sulfobutyl ether-β-cyclodextrin, the inclusion temperature, the inclusion time, the amount of ethanol, the ratio of auxiliary drugs and the stirring speed is the core technology of the present application.

[0048] The present application has the following advantages:

[0049] 1. The safety of the nimodipine for injection of the present application is significantly improved, the toxicity problem caused by the use of a large amount of organic solvent is avoided, the irritability of the preparation is reduced, and the patient's compliance is improved.

[0050] 2. The dilution stability is improved. The nimodipine for injection can be directly dispersed in 0.9% sodium chloride injection or 5% glucose injection after reconstitution, without the need for a three-way valve, thereby improving the convenience of use.

[0051] 3. The relative proportion of sulfobutyl ether-β-cyclodextrin and nimodipine in the nimodipine for injection of the present application is smaller, which not only reduces the production cost, but also reduces the safety hidden danger that may be caused by cyclodextrin.

[0052] 4. The production convenience is improved. Compared with the commonly used shaking method, the preparation time can be greatly shortened by optimizing the preparation process of the present application; at the same time, the use of a small amount of ethanol not only shortens the inclusion time, but also does not need special process to remove the small amount of ethanol in the formula, and the ethanol residue in the final product is very low, thereby increasing the production convenience and shortening the production cycle.

[0053] In summary, the present application provides a sulfobutyl ether-β-cyclodextrin-containing nimodipine cyclodextrin lyophilized powder injection, which can significantly improve the solubility, safety and dilution stability of nimodipine, increase the compliance of patients and the convenience of clinical use. Compared with the commercially available nimodipine injection, the present application has significantly reduced toxicity, significantly improved safety, dilution stability and clinical use convenience, and has obvious advantages. Compared with the existing technology of nimodipine cyclodextrin inclusion, the present application avoids the use of a large amount of organic solvents and surfactants, greatly reduces the auxiliary drug ratio, shortens the production cycle, and has very good application prospect. BRIEF DESCRIPTION OF DRAWINGS

[0054] Figure 1 . HPLC chromatogram of content determination test sample solution;

[0055] Figure 2 . Hemolysis diagram of nimodipine injection;

[0056] Figure 3 . HPLC chromatogram of related substance determination test sample solution;

[0057] Figure 4 . Dilution stability diagram of nimodipine injection and nimodipine injection;

[0058] Figure 5 . Pathological section diagram of vascular irritation of nimodipine injection;

[0059] Figure 6 . Hemolysis diagram of nimodipine injection;

[0060] Figure 7 . Pharmacodynamic evaluation diagram of nimodipine injection;

[0061] Figure 8 . Blood drug concentration-time curve diagram of nimodipine injection. DETAILED DESCRIPTION

[0062] The specific embodiments provided by the present application will be described in detail below in conjunction with the examples. It should be understood that the following examples are only used to illustrate the present application and not to limit the scope of the present application.

[0063] Example 1: Effect of polyethylene glycol 400 on nimodipine lyophilized powder injection

[0064] Referring to the operation experiment of Example 1 of patent document CN1634050A, nimodipine lyophilized powder injection was prepared.

[0065] 1. Prescription

[0066]

[0067] 2. Preparation process

[0068] The prescribed amount of nimodipine is added to a mixture of polyethylene glycol 400 and Tween-80, heated and stirred to dissolve. The hydroxypropyl-β-cyclodextrin is added to water for injection, heated and stirred to dissolve, and the above solution is added while stirring, and stirring is continued until room temperature. An appropriate amount of activated carbon is added, incubated at 60°C for 20 min and stirred. Filter, fill, freeze-drying, and you're done.

[0069] 3. Experimental results

[0070] The lyophilized powder prepared from this formulation is loose, with a shriveled, collapsed and unsatisfactory appearance, which affects the quality of the product.

[0071] Example 2: Preparation of nimodipine hydroxypropyl-β-cyclodextrin inclusion complex

[0072] Refer to the operation experiment of Example 2 of Chinese patent document CN1424035A to prepare nimodipine hydroxypropyl-β-cyclodextrin inclusion complex lyophilized powder.

[0073] 1. Prescription

[0074]

[0075] 2. Preparation process

[0076] Take 2 mg of nimodipine and stir in 2 mL of absolute ethanol until completely dissolved. Take 20 mg of hydroxypropyl-β-cyclodextrin and dissolve in 7 mL of water for injection. Add the nimodipine ethanol solution to the hydroxypropyl-β-cyclodextrin water solution while stirring to form a uniform cyclodextrin inclusion complex. Dry under reduced pressure on a rotary evaporator to remove ethanol, add 300 mg of mannitol, filter the solution through a previously sterilized vertical funnel, and place the filtrate in a sterile vial. Determine the content by high performance liquid chromatography, freeze-dry, and seal to obtain nimodipine powder for injection.

[0077] 3. The nimodipine content determination method in this invention is determined by high performance liquid chromatography (Chinese Pharmacopoeia 2020 Edition Part IV General 0512), and the operation is carried out in the dark.

[0078] Chromatographic conditions and system suitability test: use octadecylsilane bonded silica gel as the filler (C 18 Column 250 x 4.6 mm, 5 μm); with methanol-acetonitrile-water (35:38:27) as the mobile phase; detection wavelength 235 nm; injection volume 10 μL. The theoretical plate number calculated by nimodipine peak should not be less than 8000, and the separation degree of nimodipine peak from adjacent impurity peaks should meet the requirements.

[0079] Preparation of test solution: precisely take 1 mL of prepared solution, put it in a 50 mL volumetric flask, dilute to the mark with mobile phase, and shake well. The HPLC chromatogram of the test solution is shown in Figure 1

[0080] Preparation of reference solution: take nimodipine reference substance, accurately weigh, dissolve and quantitatively dilute with mobile phase to prepare a solution containing about 20 μg per 1 mL.

[0081] Determination method: accurately take the test solution and the reference solution, inject them into the liquid chromatograph respectively, and record the chromatogram. Calculate by the external standard method with peak area.

[0082] 4. Experimental results

[0083] During the mixing and stirring of the nimodipine ethanol solution and the hydroxypropyl-β-cyclodextrin aqueous solution, the drug precipitated. After filtering the sample, the drug content in the solution was 0.005 mg / mL, and the auxiliary drug ratio was calculated to be 571.

[0084] 5. Analysis of results

[0085] The use of 22% ethanol in this patent technology is not conducive to the inclusion of nimodipine by cyclodextrin, and a large amount of ethanol poses a certain safety hazard; in addition, the removal of ethanol by rotary evaporation increases the complexity of the process.

[0086] Example 3: Preparation of nimodipine hydroxypropyl-β-cyclodextrin inclusion complex

[0087] Refer to the operation experiment of Example 28 of Chinese patent document CN1653089A to prepare nimodipine hydroxypropyl-β-cyclodextrin inclusion complex.

[0088] 1. Prescription

[0089]

[0090] 2. Preparation process

[0091] Dissolve 0.5 g of nimodipine in the prescribed amount of ethanol, weigh 68 g of hydroxypropyl-β-cyclodextrin in an appropriate amount of water, mix the above solutions, stir and heat at 60°C, add activated carbon to remove heat sources, filter, input the filtrate into a 1 liter rotary tank, keep warm at 70-80°C, reduce pressure to concentrate, recover the solvent and convert it to a water system, continue to keep warm, reduce pressure to swell the material, and dry.

[0092] 3. Experimental results

[0093] ​After repeated verification by experiments, the drug was precipitated during the mixing and stirring of the nimodipine ethanol solution and the hydroxypropyl-β-cyclodextrin aqueous solution; the drug was also precipitated in large quantities during the removal of ethanol by reduced pressure drying. After filtration of the sample, the drug content in the solution was 0.21 mg / mL, and the adjuvant ratio was calculated to be 1904, which was much higher than the adjuvant ratio of the application.

[0094] Example 4: Preparation of nimodipine sulfobutyl ether-β-cyclodextrin inclusion complex by shaking method

[0095] The shaking method was used to prepare the nimodipine sulfobutyl ether-β-cyclodextrin inclusion complex according to the literature (Maria DN, et al. Nimodipine Ophthalmic Formulations for Management of Glaucoma. Pharm Res. 2017; 34(4): 809-824.), and in addition, the same formula containing a small amount of ethanol was used to prepare the nimodipine sulfobutyl ether-β-cyclodextrin inclusion complex by shaking method for comparison.

[0096] 1. Prescription

[0097] Table 1 Prescription design

[0098] Ingredients Formulation I Formulation II Nimodipine 30 mg 30 mg Ethanol - 0.6 mL Sulfobutyl ether-β-cyclodextrin 1.3g 1.3g Water for injection q.s. 30 mL q.s. 30 mL

[0099] 2. Preparation process

[0100] Take sulfobutyl ether-β-cyclodextrin 1.3 g, in parallel in two groups, add an appropriate amount of water for injection, stir to dissolve, and obtain a sulfobutyl ether-β-cyclodextrin aqueous solution; one group does not add ethanol, and the other group takes the prescribed amount of ethanol and adds it to the sulfobutyl ether-β-cyclodextrin aqueous solution, and two groups are respectively added with water for injection to make up to 30 mL, placed in a Westlin bottle, added with the prescribed amount of nimodipine, plugged with a stopper, the suspension was vortexed for 5 min, then ultrasonic for 15 min, and oscillated in a 37°C electric heating constant temperature water bath for 72 h. The obtained solution was filtered through a 0.45 μm polyether sulfone filter membrane, and the content of nimodipine was detected by HPLC method.

[0101] 3. Experimental results

[0102] Table 2 Solubility of nimodipine sulfobutyl ether-β-cyclodextrin inclusion complex prepared by shaking method

[0103] Inspection items Formulation I Formulation II Drug content (mg / mL) 0.10 0.09 Ratio of adjuvant 456 462

[0104] The results show that even if a small amount of ethanol is added to assist dissolution in the prescription, the nimodipine sulfobutyl ether-β-cyclodextrin inclusion complex prepared by shaking for 72 h has poor inclusion effect, and the preparation time is long, which is not conducive to the industrialized production of injections.

[0105] Example 5: Effect of different solvents on solubility of nimodipine in sulfobutyl ether-β-cyclodextrin

[0106] 1. Formulation

[0107] Table 3 Formulation design

[0108]

[0109]

[0110] 2. Preparation process

[0111] Take sulfobutyl ether-β-cyclodextrin 9 g, parallel 5 groups, add appropriate amount of water for injection, stir to dissolve at 30°C, get sulfobutyl ether-β-cyclodextrin aqueous solution; take nimodipine 45 mg, add 0.9 mL of solvent to dissolve, under stirring conditions, add nimodipine solution to sulfobutyl ether-β-cyclodextrin aqueous solution, stir for 2 h, filter the obtained solution through 0.45 μm polyether sulfone filter membrane, detect the content of nimodipine by HPLC method.

[0112] 3. Experimental results

[0113] Table 4 Effect of different solvents on solubility of nimodipine in sulfobutyl ether-β-cyclodextrin

[0114]

[0115] The results show that adding an appropriate amount of organic solvent is beneficial to the solubilization of nimodipine by cyclodextrin, and the best effect is ethanol.

[0116] Example 6: Effect of ethanol dosage on solubility of nimodipine in sulfobutyl ether-β-cyclodextrin

[0117] 1. Formulation

[0118] Table 5 Formulation design

[0119]

[0120] 2. Preparation process

[0121] Take sulfobutyl ether-β-cyclodextrin 6 g, parallel 7 groups, add appropriate amount of water for injection, stir to dissolve at 30°C, get sulfobutyl ether-β-cyclodextrin aqueous solution; take nimodipine 15 mg, respectively add 0.6 mL, 0.9 mL, 1.2 mL, 1.5 mL, 1.8 mL of ethanol to dissolve, under stirring conditions, add nimodipine ethanol solution to sulfobutyl ether-β-cyclodextrin aqueous solution, stir for 2 h, filter the obtained solution through 0.45 μm polyether sulfone filter membrane, detect the content of nimodipine by HPLC method.

[0122] 3. Experimental results

[0123] The content of the prepared nimodipine cyclodextrin was determined, and the results were as follows:

[0124] Table 6 Effect of different ethanol amounts on solubility of nimodipine in sulfobutyl ether-β-cyclodextrin

[0125]

[0126] The experimental results show that with the increase of ethanol amount, the solubility of nimodipine in sulfobutyl ether-β-cyclodextrin aqueous solution decreases, and the auxiliary drug ratio increases.

[0127] Example 7: Effect of sulfobutyl ether-β-cyclodextrin amount on solubility of nimodipine

[0128] 1. Prescription

[0129] Table 7 Prescription design

[0130]

[0131] 2. Preparation process

[0132] Take 3g, 6g, 7.5g, 9g, 10.5g, 12g, 13.5g, 15g of sulfobutyl ether-β-cyclodextrin, add appropriate amount of water for injection, stir to dissolve at 30°C, get sulfobutyl ether-β-cyclodextrin aqueous solution; take 45mg of nimodipine, dissolve in 0.9mL of ethanol, under stirring conditions, add the nimodipine ethanol solution to the sulfobutyl ether-β-cyclodextrin aqueous solution, stir for 2h, the obtained solution is filtered through a 0.45μm polyether sulfone filter membrane, and the content of nimodipine is detected by HPLC method.

[0133] 3. Experimental results

[0134] The content of the prepared nimodipine cyclodextrin was determined, and the results were as follows:

[0135] Table 8 Effect of different sulfobutyl ether-β-cyclodextrin amounts on solubility of nimodipine

[0136]

[0137] The experimental results show that when the amount of sulfobutyl ether-β-cyclodextrin is less than 45%, with the increase of the amount of sulfobutyl ether-β-cyclodextrin, the solubility of nimodipine increases, and the auxiliary drug ratio significantly decreases; and when the amount of sulfobutyl ether-β-cyclodextrin is greater than 45%, the solubility of nimodipine does not significantly increase, and the auxiliary drug ratio increases.

[0138] Example 8: Effect of dissolution temperature and time on solubility of nimodipine

[0139] 1. Prescription

[0140] Table 9 Prescription design

[0141]

[0142] 2. Preparation process

[0143] Take sulfobutyl ether-β-cyclodextrin 12 g, 9 groups in parallel, add an appropriate amount of water for injection, stir to dissolve, get sulfobutyl ether-β-cyclodextrin aqueous solution; take nimodipine 60 mg respectively, add 1.2 mL of ethanol to dissolve, under stirring conditions, nimodipine ethanol solution is added to sulfobutyl ether-β-cyclodextrin aqueous solution solution, respectively at 20℃, 30℃, 40℃, 50℃, 60℃, 70℃, 80℃, 90℃, 100℃ stirring 5h, 1h, 2h, 3h, 4h, 5h take point, the obtained solution is filtered by 0.45μm polyether sulfone filter membrane, HPLC method is used to detect the content of nimodipine.

[0144] 3. Results analysis

[0145] The content of nimodipine cyclodextrin taken at different times is determined, and the results are as follows

[0146] Table 10 Solubility of nimodipine stirred for 5h at different temperatures

[0147]

[0148] Table 11 Sulfobutyl ether-β-cyclodextrin and nimodipine ratio stirred for 5h at different temperatures

[0149]

[0150] Experiments show that as the temperature increases, the solubility of nimodipine increases, and the ratio of adjuvant decreases; under the condition of lower temperature, as the stirring time increases, the solubility of nimodipine decreases, and the ratio of adjuvant increases. This shows that increasing the temperature is beneficial to the inclusion of sulfobutyl ether-β-cyclodextrin to nimodipine, but too high temperature may lead to the generation of related substances.

[0151] Example 9: Preparation of nimodipine for injection

[0152] 1. Prescription

[0153]

[0154] 2. Preparation process

[0155] Take sulfobutyl ether-β-cyclodextrin 3g, add the appropriate amount of water for injection, stirring to dissolve at 20°C, sulfobutyl ether-β-cyclodextrin aqueous solution; take nimodipine 3mg, dissolved in 0.06mL ethanol, nimodipine ethanol solution under stirring conditions into the sulfobutyl ether-β-cyclodextrin aqueous solution solution, stirring 10min, add water for injection to 30mL, the resulting solution was sterile filtered through 0.22μm microporous filter, filling, freeze-drying, nitrogen, packaging, ready. The whole process of dissolution and filtration into clean dry N2.

[0156] Example 10: preparation of nimodipine for injection

[0157] 1. prescription

[0158]

[0159] 2. preparation process

[0160] Take sulfobutyl ether-β-cyclodextrin 6g, add the appropriate amount of water for injection, stirring to dissolve at 40°C, sulfobutyl ether-β-cyclodextrin aqueous solution; take nimodipine 15mg, dissolved in 0.3mL ethanol, nimodipine ethanol solution under stirring conditions into the sulfobutyl ether-β-cyclodextrin aqueous solution solution, stirring 30min, add water for injection to 30mL, the resulting solution was sterile filtered through 0.22μm microporous filter, filling, freeze-drying, packaging, ready.

[0161] Example 11: preparation of nimodipine for injection

[0162] 1. prescription

[0163]

[0164] 2. preparation process

[0165] Take sulfobutyl ether-β-cyclodextrin 10.5g, sodium citrate 60mg and citric acid 9mg, add the appropriate amount of water for injection, stirring to dissolve at 60°C, sulfobutyl ether-β-cyclodextrin aqueous solution; take nimodipine 15mg, under stirring conditions into the sulfobutyl ether-β-cyclodextrin aqueous solution solution, stirring 2h, add water for injection to 30mL, the resulting solution was sterile filtered through 0.22μm microporous filter, filling, freeze-drying, nitrogen, packaging, ready. The whole process of dissolution and filtration into clean dry N2.

[0166] Example 12: preparation of nimodipine for injection

[0167] 1. prescription

[0168]

[0169]

[0170] 2. Preparation process

[0171] Take sulfobutyl ether-β-cyclodextrin 9g, add an appropriate amount of water for injection, 50°C stirring to dissolve, get sulfobutyl ether-β-cyclodextrin aqueous solution; take nimodipine 24mg, dissolved in 0.48ml ethanol, under stirring conditions, nimodipine ethanol solution is added to the sulfobutyl ether-β-cyclodextrin aqueous solution solution, stirring 1h, add water for injection to 30ml, the resulting solution is sterile filtered by 0.22μm microporous filter, filling, freeze-drying, nitrogen, packaging, ready. The whole process of dissolution and filtration into clean dry N2.

[0172] Example 13: preparation of nimodipine for injection

[0173] 1. Prescription

[0174]

[0175] 2. Preparation process

[0176] Take sulfobutyl ether-β-cyclodextrin 10.8g, add an appropriate amount of water for injection, 60°C stirring to dissolve, get sulfobutyl ether-β-cyclodextrin aqueous solution; take nimodipine 24mg, dissolved in 0.3ml ethanol, under stirring conditions, nimodipine ethanol solution is added to the sulfobutyl ether-β-cyclodextrin aqueous solution solution, stirring 1h, add water for injection to 30ml, the resulting solution is sterile filtered by 0.22μm microporous filter, filling, freeze-drying, packaging, ready.

[0177] Example 14: preparation of nimodipine for injection

[0178] 1. Prescription

[0179]

[0180] 2. Preparation process

[0181] Take sulfobutyl ether-β-cyclodextrin 12g, disodium hydrogen phosphate 709mg and citric acid 102mg, add an appropriate amount of water for injection, 70°C stirring to dissolve, get sulfobutyl ether-β-cyclodextrin aqueous solution; take nimodipine 24mg, under stirring conditions, nimodipine is added to the sulfobutyl ether-β-cyclodextrin aqueous solution solution, stirring 2h, add water for injection to 30ml, the resulting solution is sterile filtered by 0.22μm microporous filter, filling, freeze-drying, packaging, ready.

[0182] Example 15: preparation of nimodipine for injection

[0183] 1. Prescription

[0184]

[0185] 2. Preparation process

[0186] Take sulfobutyl ether-β-cyclodextrin 9 g, add an appropriate amount of water for injection, 50 ℃ stirring to dissolve, get sulfobutyl ether-β-cyclodextrin aqueous solution; take nimodipine 30 mg, dissolved in 0.3 mL of ethanol, under stirring conditions, the nimodipine ethanol solution is added to the sulfobutyl ether-β-cyclodextrin aqueous solution solution, stirring 2 h, add water for injection to 30 mL, the resulting solution is sterile filtered through 0.22 μm microporous filter, filling, freeze-drying, nitrogen, packaging, ready. The whole process of dissolution and filtration into clean dry N2.

[0187] Example 16: Preparation of nimodipine for injection

[0188] 1. Prescription

[0189]

[0190] 2. Preparation process

[0191] Take sulfobutyl ether-β-cyclodextrin 9.9 g, disodium hydrogen phosphate 520 mg and sodium dihydrogen phosphate 281 mg, add an appropriate amount of water for injection, 60 ℃ stirring to dissolve, get sulfobutyl ether-β-cyclodextrin aqueous solution; take nimodipine 30 mg, dissolved in 0.3 mL of ethanol, under stirring conditions, the nimodipine ethanol solution is added to the sulfobutyl ether-β-cyclodextrin aqueous solution solution, stirring 2 h, add water for injection to 30 mL, the resulting solution is sterile filtered through 0.22 μm microporous filter, filling, freeze-drying, packaging, ready.

[0192] Example 17: Preparation of nimodipine for injection

[0193] 1. Prescription

[0194]

[0195] 2. Preparation process

[0196] Take sulfobutyl ether-β-cyclodextrin 11.1 g, add an appropriate amount of water for injection, 60 ℃ stirring to dissolve, get sulfobutyl ether-β-cyclodextrin aqueous solution; take nimodipine 30 mg, dissolved in 0.48 mL of ethanol, under stirring conditions, the nimodipine ethanol solution is added to the sulfobutyl ether-β-cyclodextrin aqueous solution solution, stirring 2 h, add water for injection to 30 mL, the resulting solution is sterile filtered through 0.22 μm microporous filter, filling, freeze-drying, nitrogen, packaging, ready. The whole process of dissolution and filtration into clean dry N2.

[0197] Example 18: Preparation of nimodipine for injection

[0198] 1. Formulation

[0199]

[0200] 2. Preparation process

[0201] Take sulfobutyl ether-β-cyclodextrin 9 g, add an appropriate amount of water for injection, 60 ℃ stirring to dissolve, get sulfobutyl ether-β-cyclodextrin aqueous solution; take nimodipine 30 mg, dissolved in 0.48 mL of ethanol, under stirring conditions, the nimodipine ethanol solution is added to the sulfobutyl ether-β-cyclodextrin aqueous solution solution, stirring 3 h, add water for injection to 30 mL, the resulting solution is sterile filtered through 0.22 μm microporous filter, fill, freeze-drying, nitrogen, packaging, get. The whole process of dissolution and filtration into clean dry N2.

[0202] Example 19: Preparation of nimodipine for injection

[0203] 1. Formulation

[0204]

[0205]

[0206] 2. Preparation process

[0207] Take sulfobutyl ether-β-cyclodextrin 9 g, add an appropriate amount of water for injection, 100 ℃ stirring to dissolve, get sulfobutyl ether-β-cyclodextrin aqueous solution; take nimodipine 30 mg, dissolved in 0.72 mL of ethanol, under stirring conditions, the nimodipine ethanol solution is added to the sulfobutyl ether-β-cyclodextrin aqueous solution solution, stirring 2 h, add water for injection to 30 mL, the resulting solution is sterile filtered through 0.22 μm microporous filter, fill, freeze-drying, packaging, get.

[0208] Example 20: Preparation of nimodipine for injection

[0209] 1. Formulation

[0210]

[0211] 2. Preparation process

[0212] Take sulfobutyl ether-β-cyclodextrin 10.5 g, add the appropriate amount of water for injection, 50 ℃ stirring to dissolve, get sulfobutyl ether-β-cyclodextrin aqueous solution; take nimodipine 30 mg, dissolved in 0.48 mL ethanol, under stirring conditions, the nimodipine ethanol solution is added to the sulfobutyl ether-β-cyclodextrin aqueous solution solution, stirring 5 h, add water for injection to 30 mL, the resulting solution is sterile filtered through 0.22 μm microporous filter, filling, freeze-drying, packaging, get.

[0213] Example 21: Preparation of nimodipine for injection

[0214] 1. Prescription

[0215]

[0216] 2. Preparation process

[0217] Take sulfobutyl ether-β-cyclodextrin 10.5 g, add the appropriate amount of water for injection, 60 ℃ stirring to dissolve, get sulfobutyl ether-β-cyclodextrin aqueous solution; take nimodipine 30 mg, dissolved in 0.72 mL ethanol, under stirring conditions, the nimodipine ethanol solution is added to the sulfobutyl ether-β-cyclodextrin aqueous solution solution, stirring 1 h, add water for injection to 30 mL, the resulting solution is sterile filtered through 0.22 μm microporous filter, filling, freeze-drying, packaging, get.

[0218] Example 22: Preparation of nimodipine for injection

[0219] 1. Prescription

[0220]

[0221] 2. Preparation process

[0222] Take sulfobutyl ether-β-cyclodextrin 10.5 g, add the appropriate amount of water for injection, 70 ℃ stirring to dissolve, get sulfobutyl ether-β-cyclodextrin aqueous solution; take nimodipine 30 mg, dissolved in 0.9 mL ethanol, under stirring conditions, the nimodipine ethanol solution is added to the sulfobutyl ether-β-cyclodextrin aqueous solution solution, stirring 2 h, add water for injection to 30 mL, the resulting solution is sterile filtered through 0.22 μm microporous filter, filling, freeze-drying, nitrogen, packaging, get. The whole process of dissolution and filtration into clean dry N2.

[0223] Example 23: Preparation of nimodipine for injection

[0224] 1. Prescription

[0225]

[0226] 2. Preparation process

[0227] Take sulfobutyl ether-β-cyclodextrin 10.5 g, add an appropriate amount of water for injection, 80°C stirring to dissolve, get sulfobutyl ether-β-cyclodextrin aqueous solution; take nimodipine 30 mg, under stirring conditions nimodipine is added to the sulfobutyl ether-β-cyclodextrin aqueous solution solution, stirring 3 h, add water for injection to 30 mL, the resulting solution is sterile filtered through 0.22 μm microporous filter, fill, freeze-drying, nitrogen, packaging, ready. The whole process of dissolution and filtration into clean dry N2.

[0228] Example 24: preparation of nimodipine for injection

[0229] 1. prescription

[0230]

[0231]

[0232] 2. preparation process

[0233] Take sulfobutyl ether-β-cyclodextrin 12 g, add an appropriate amount of water for injection, 60°C stirring to dissolve, get sulfobutyl ether-β-cyclodextrin aqueous solution; take nimodipine 30 mg, dissolved in 0.9 mL ethanol, under stirring conditions nimodipine ethanol solution is added to the sulfobutyl ether-β-cyclodextrin aqueous solution solution, stirring 2 h, add water for injection to 30 mL, the resulting solution is sterile filtered through 0.22 μm microporous filter, fill, freeze-drying, packaging, ready.

[0234] Example 25: preparation of nimodipine for injection

[0235] 1. prescription

[0236]

[0237] 2. preparation process

[0238] Take sulfobutyl ether-β-cyclodextrin 12 g, disodium hydrogen phosphate 520 mg, sodium dihydrogen phosphate 281 mg, add an appropriate amount of water for injection, 60°C stirring to dissolve, get sulfobutyl ether-β-cyclodextrin aqueous solution; take nimodipine 30 mg, under stirring conditions nimodipine is added to the sulfobutyl ether-β-cyclodextrin aqueous solution solution, stirring 2 h, add water for injection to 30 mL, the resulting solution is sterile filtered through 0.22 μm microporous filter, fill, freeze-drying, packaging, ready.

[0239] Example 26: preparation of nimodipine for injection

[0240] 1. prescription

[0241]

[0242] 2. Preparation process

[0243] Take sulfobutyl ether-β-cyclodextrin 13.5 g, add an appropriate amount of water for injection, 50°C stirring to dissolve, get sulfobutyl ether-β-cyclodextrin aqueous solution; take nimodipine 30 mg, dissolved in 0.48 mL of ethanol, under stirring conditions, the nimodipine ethanol solution is added to the sulfobutyl ether-β-cyclodextrin aqueous solution solution stirring 3 h, add water for injection to 30 mL, the resulting solution is sterile filtered through 0.22 μm microporous filter, filling, freeze-drying, packaging, ready.

[0244] Example 27: Preparation of nimodipine for injection

[0245] 1. Prescription

[0246]

[0247] 2. Preparation process

[0248] Take sulfobutyl ether-β-cyclodextrin 12 g, add an appropriate amount of water for injection, 70°C stirring to dissolve, get sulfobutyl ether-β-cyclodextrin aqueous solution; take nimodipine 36 mg, dissolved in 0.72 mL of ethanol, under stirring conditions, the nimodipine ethanol solution is added to the sulfobutyl ether-β-cyclodextrin aqueous solution solution stirring 3 h, add water for injection to 30 mL, the resulting solution is sterile filtered through 0.22 μm microporous filter, filling, freeze-drying, packaging, ready.

[0249] Example 28: Preparation of nimodipine for injection

[0250] 1. Prescription

[0251]

[0252] 2. Preparation process

[0253] Take sulfobutyl ether-β-cyclodextrin 12.6 g, disodium hydrogen phosphate 59 mg and potassium dihydrogen phosphate 6.7 mg, add an appropriate amount of water for injection, 60°C stirring to dissolve, get sulfobutyl ether-β-cyclodextrin aqueous solution; take nimodipine 36 mg, dissolved in 0.72 mL of ethanol, under stirring conditions, the nimodipine ethanol solution is added to the sulfobutyl ether-β-cyclodextrin aqueous solution solution stirring 2 h, add water for injection to 30 mL, the resulting solution is sterile filtered through 0.22 μm microporous filter, filling, freeze-drying, packaging, ready.

[0254] Example 29: Preparation of nimodipine for injection

[0255] 1. Prescription

[0256]

[0257] 2. Preparation process

[0258] Take sulfobutyl ether-β-cyclodextrin 13.5 g, add an appropriate amount of water for injection, 100 ℃ stirring to dissolve, get sulfobutyl ether-β-cyclodextrin aqueous solution; take nimodipine 36 mg, under stirring conditions nimodipine is added to the sulfobutyl ether-β-cyclodextrin aqueous solution solution, stirring 1 h, add water for injection to 30 mL, the resulting solution is sterile filtered through 0.22 μm microporous filter, filling, freeze-drying, nitrogen, packaging, ready. The whole process of dissolution and filtration into clean dry N2.

[0259] Example 30: preparation of nimodipine for injection

[0260] 1. Prescription

[0261]

[0262] 2. Preparation process

[0263] Take sulfobutyl ether-β-cyclodextrin 13.5 g, add an appropriate amount of water for injection, 80 ℃ stirring to dissolve, get sulfobutyl ether-β-cyclodextrin aqueous solution; take nimodipine 45 mg, dissolved in 0.9 mL ethanol, under stirring conditions nimodipine ethanol solution is added to the sulfobutyl ether-β-cyclodextrin aqueous solution solution, stirring 4 h, add water for injection to 30 mL, the resulting solution is sterile filtered through 0.22 μm microporous filter, filling, freeze-drying, packaging, ready.

[0264] Example 31: preparation of nimodipine for injection

[0265] 1. Prescription

[0266]

[0267] 2. Preparation process

[0268] Take sulfobutyl ether-β-cyclodextrin 15 g, add an appropriate amount of water for injection, 100 ℃ stirring to dissolve, get sulfobutyl ether-β-cyclodextrin aqueous solution; take nimodipine 60 mg, dissolved in 1.5 mL ethanol, under stirring conditions nimodipine ethanol solution is added to the sulfobutyl ether-β-cyclodextrin aqueous solution solution, stirring 5 h, add water for injection to 30 mL, the resulting solution is sterile filtered through 0.22 μm microporous filter, filling, freeze-drying, nitrogen, packaging, ready. The whole process of dissolution and filtration into clean dry N2.

[0269] Example 32: preparation of nimodipine for injection

[0270] 1. Prescription

[0271]

[0272] 2. Preparation process

[0273] Take sulfobutyl ether-β-cyclodextrin 12 g, potassium dihydrogen phosphate 204 mg and sodium hydroxide 3.5 mg, add an appropriate amount of water for injection, stir to dissolve at 100 ℃, and obtain a sulfobutyl ether-β-cyclodextrin aqueous solution; take nimodipine 60 mg, dissolve in 0.9 mL of ethanol, and add the nimodipine ethanol solution to the sulfobutyl ether-β-cyclodextrin aqueous solution under stirring for 5 h. Add water for injection to make up to 30 mL. The obtained solution is sterile filtered through a 0.22 μm microporous filter, filled, freeze-dried, and packaged.

[0274] Example 33: Ethanol residue determination of nimodipine for injection

[0275] Select nimodipine for injection prepared in examples 9, 10, 12, 13, 15, 16, 17, 18, 19, 20, 21, 22, 24, 26, 27, 28, 30, 31, 32, and detect the ethanol residue.

[0276] 1. The ethanol content in nimodipine for injection is determined by residual solvent determination method (Chinese Pharmacopoeia 2020 edition four general rules 0861 second method) as follows:

[0277] Chromatographic conditions:

[0278] Use DB-624 capillary column as chromatographic column; program temperature, initial temperature is 70 ℃, maintain for 2 min, first increase the temperature to 120 ℃ at a rate of 10 ℃ per min, then increase the temperature to 220 ℃ at a rate of 20 ℃ per min, maintain for 3 min; split ratio is 20:1; column flow is 1 mL / min; injection port temperature is 200 ℃; detector temperature is 250 ℃; headspace bottle equilibrium temperature is 80 ℃, equilibrium time is 30 min.

[0279] Preparation of blank solution:

[0280] Precisely weigh 2 g of prepared blank auxiliary material freeze-dried powder into three 20 mL volumetric flasks, dilute with DMF to the mark, shake well, and accurately take 4 mL into a headspace bottle, seal, and obtain.

[0281] Preparation of reference solution:

[0282] Take an appropriate amount of ethanol, accurately weigh, and dilute with DMF to prepare a solution containing 5 mg per 1 mL as a reference stock solution. Accurately take 2 mL of the reference stock solution into a 20 mL volumetric flask, dilute with DMF to the mark, shake well, accurately take 4 mL into a headspace bottle, seal, and obtain the reference solution.

[0283] Preparation of test sample solution:

[0284] Precisely weigh 2g of nimodipine for injection, put it into three 20mL volumetric flasks, dilute to the scale with DMF, shake well, precisely take 4mL, put it into the empty bottle, seal, and the test sample solution is obtained.

[0285] Determination method:

[0286] Take the blank solution and the control sample solution into the headspace, record the chromatogram. Take the test sample solution and the control sample solution into the headspace respectively, record the chromatogram, and calculate by the external standard method with the peak area.

[0287] 2. Experimental results

[0288] Table 12 Nimodipine for injection ethanol residue results

[0289] Sample name Ethanol residue (%) Nimodipine for injection (Example 9) 0.16 Nimodipine for injection (Example 10) 0.29 Nimodipine for injection (Example 12) 0.24 Nimodipine for injection (Example 13) 0.02 Nimodipine for injection (Example 15) 0.12 Nimodipine for injection (Example 16) 0.02 Nimodipine for injection (Example 17) 0.03 Nimodipine for injection (Example 18) 0.02 Nimodipine for injection (Example 19) 0.01 Nimodipine for injection (Example 20) 0.03 Nimodipine for injection (Example 21) 0.08 Nimodipine for injection (Example 22) 0.06 Nimodipine for injection (Example 24) 0.04 Nimodipine for injection (Example 26) 0.07 Nimodipine for injection (Example 27) 0.03 Nimodipine for injection (Example 28) 0.04 Nimodipine for injection (Example 30) 0.02 Nimodipine for injection (Example 31) 0.03 Nimodipine for injection (Example 32) 0.02

[0290] The results show that by controlling the temperature and time, the ethanol residue of nimodipine for injection of the application meets the quality standard requirements.

[0291] Example 34: Dilution stability study of nimodipine for injection

[0292] 1. Experimental method

[0293] Take the nimodipine for injection prepared in Example 21 after reconstitution and the nimodipine injection, respectively, dilute to the clinical administration concentration with 0.9% sodium chloride injection and 5% glucose. At 0, 3, 6, 9, 12, 24h, observe and take samples, filter with 0.45μm polyether sulfone filter membrane, and determine the nimodipine content according to the above content determination method.

[0294] 2. Experimental results

[0295] Table 13 Content change of nimodipine for injection and nimodipine injection diluted with 0.9% sodium chloride injection

[0296]

[0297] Note: - indicates no precipitation, + indicates precipitation.

[0298] Table 14 Content change of nimodipine for injection and nimodipine injection diluted with 5% glucose

[0299]

[0300]

[0301] Note: - indicates no precipitation, + indicates precipitation.

[0302] When nimodipine injection is diluted with 0.9% sodium chloride injection or 5% glucose, a large amount of the drug precipitates out within 3 hours, resulting in a significant decrease in content. After 24 hours, the contents are only 2.30% and 6.24%, respectively, indicating extremely poor dilution stability. In contrast, the nimodipine for injection of this invention, after dilution with 0.9% sodium chloride injection or 5% glucose, shows no significant change in content within 24 hours, remains clear, and exhibits good dilution stability. These results demonstrate that, compared to nimodipine injection, the nimodipine for injection of this invention has significantly improved dilution stability and exhibits a clear advantage.

[0303] Example 35: Hemolytic activity study of nimodipine for injection

[0304] 1. Experimental Methods

[0305] The nimodipine for injection prepared in Example 21 and the commercially available nimodipine injection were diluted with 0.9% sodium chloride injection to a clinically administered concentration of 0.04 mg / mL. Eight clean 10 mL glass test tubes were taken and numbered: tubes 1 to 5 contained different concentrations of nimodipine for injection, tube 6 was the negative control, tube 7 was the positive control, and tube 8 contained the commercially available nimodipine injection. As shown in the table below, 2% red blood cell suspension, 0.9% sodium chloride injection, distilled water, and the drug solution were added sequentially. After mixing, the tubes were immediately incubated in a water bath at 37±0.5℃. The hemolysis status of each tube was observed and recorded. Initially, records were taken every 15 minutes, then every hour after 1 hour, for a total of 3 hours.

[0306] Table 15 Hemolysis Experimental Design

[0307]

[0308] 2. Experimental Results

[0309] Hemolysis such as Figure 2 As shown, none of the injectable nimodipine (Nos. 1-5) of the present invention exhibited hemolysis, indicating that the injectable nimodipine of the present invention has good safety.

[0310] Example 36: High-Temperature Stability Study of Nimodipine for Injection

[0311] Nimodipine for injection prepared in Example 21 was placed at 60°C for one month, and the drug content and changes in related impurities were detected.

[0312] 1. The method for determining nimodipine-related substances in this invention is performed by high performance liquid chromatography (General Chapter 0512, Part IV, Chinese Pharmacopoeia 2020 Edition), and the operation is carried out in the dark.

[0313] Chromatographic conditions and system suitability test: Octadecylsilane-bonded silica gel was used as the stationary phase (C 18Column 250 x 4.6 mm, 5 μm); mobile phase: methanol-acetonitrile-water (35:38:27); detection wavelength: 235 nm; injection volume: 10 μL. The separation degree between the peak of nimodipine and the peaks of impurities B, C and I in the chromatogram of the system suitability solution should be greater than 3.0.

[0314] Preparation of test sample solution: accurately take the product, dilute with the mobile phase to prepare a solution containing about 0.2 mg of nimodipine per 1 mL. The HPLC chromatogram of the test sample solution for the determination of related substances is shown in Figure 3 .

[0315] Preparation of control solution: accurately take 1 mL of the test sample solution, place it in a 100 mL volumetric flask, dilute to the mark with the mobile phase, and shake well.

[0316] Preparation of impurity I control solution: accurately weigh the impurity I control, dilute with the mobile phase to prepare a solution containing about 1 μg per 1 mL.

[0317] Preparation of system suitability solution: take nimodipine and impurities B, C and I controls, dissolve and dilute with the mobile phase to prepare a mixed solution containing about 200 μg and 1 μg per 1 mL.

[0318] Determination method: accurately take the test sample solution, impurity I control solution and control solution, and inject them into the liquid chromatograph. If there are impurity peaks in the chromatogram of the test sample solution, except for the chromatographic peaks with relative retention time less than 0.45, the chromatographic peaks with retention time consistent with that of impurity I peak are calculated by the external standard method in terms of peak area; other single impurities are calculated by the self-control method of the main component.

[0319] 2. Experimental results

[0320] Table 16 Experimental results of nimodipine for injection at high temperature stability

[0321]

[0322]

[0323] 2. Analysis of results

[0324] After storing in a 60℃ stability test box for 1 month, the labeled percentage content of nimodipine for injection has almost no change, and the related substances meet the requirements of the specified limits. The above results show that the nimodipine for injection of the present application has good high temperature stability, and the quality meets the relevant requirements.

[0325] Example 37: Effect of preparation temperature and time on the solubility of nimodipine in sulfobutyl ether-β-cyclodextrin

[0326] The effects of preparation temperature and time on the solubility of nimodipine in sulfobutyl ether-β-cyclodextrin and the related substances were investigated by taking the content of nimodipine and the related substances as indexes.

[0327] 1. Formulation

[0328] Table 17 Formulation design

[0329]

[0330] 2. Preparation process

[0331] Sulfobutyl ether-β-cyclodextrin 12 g was taken in parallel in 9 groups, and an appropriate amount of water for injection was added, and stirred to dissolve, to obtain a sulfobutyl ether-β-cyclodextrin aqueous solution; nimodipine 60 mg was taken, and under the condition of a stirring speed of 2.0 m / s, the nimodipine was added to the sulfobutyl ether-β-cyclodextrin aqueous solution, and stirred at 20℃, 30℃, 40℃, 50℃, 60℃, 70℃, 80℃, 90℃, and 100℃ for 5 h, and samples were taken at 1 h, 2 h, 3 h, 4 h, and 5 h, and the obtained solutions were filtered through a 0.45 μm polyether sulfone filter membrane, and the content of nimodipine and the related substances were detected by HPLC.

[0332] 3. Experimental results

[0333] The content and related substances of nimodipine cyclodextrin inclusion were determined at different times, and the results were as follows.

[0334] Table 18 Solubility of nimodipine stirred for 5 h in sulfobutyl ether-β-cyclodextrin at different temperatures

[0335]

[0336]

[0337] Table 19 Ratio of sulfobutyl ether-β-cyclodextrin and nimodipine stirred for 5 h at different temperatures

[0338]

[0339] Table 20 Related substances of nimodipine cyclodextrin stirred for 5 h at different temperatures

[0340]

[0341]

[0342] The experiments showed that without the addition of ethanol, the solubility of nimodipine increased with the increase of temperature and stirring time, and the ratio of adjuvant decreased; the related substances of nimodipine increased with the increase of temperature and stirring time. Therefore, appropriate preparation temperature and time should be selected for the preparation of nimodipine sulfobutyl ether-β-cyclodextrin inclusion.

[0343] Example 38: Effect of stirring speed on solubility of nimodipine in sulfobutyl ether-β-cyclodextrin

[0344] 1. Formulation

[0345] Table 21 Formulation design

[0346]

[0347] 2. Preparation process

[0348] Take sulfobutyl ether-β-cyclodextrin 25 g, 8 groups in parallel, add an appropriate amount of water for injection, stir at 75°C to dissolve, get sulfobutyl ether-β-cyclodextrin aqueous solution; take nimodipine 50 mg, respectively, under the conditions of stirring speed of 0.25 m / s, 0.5 m / s, 1.0 m / s, 1.5 m / s, 2.0 m / s, 3.0 m / s, 6.0 m / s, 10.0 m / s, nimodipine is added to sulfobutyl ether-β-cyclodextrin aqueous solution, stirring for 1.5 h, adding water for injection to 100 mL, the obtained solution is filtered by 0.45 μm polyether sulfone filter membrane, and the content of nimodipine is detected by HPLC method.

[0349] 3. Experimental results

[0350] The effect of stirring speed on the solubility of nimodipine in sulfobutyl ether-β-cyclodextrin is shown in Table 22. The results show that the drug content increases with the increase of stirring speed, and the stirring speed is too low to achieve the ideal drug loading concentration.

[0351] Table 22 Effect of stirring speed on solubility of nimodipine in sulfobutyl ether-β-cyclodextrin

[0352]

[0353] 4. Discussion

[0354] The study shows that the stirring speed significantly affects the inclusion effect of sulfobutyl ether-β-cyclodextrin on nimodipine, the greater the stirring speed, the better the inclusion effect, and the higher the drug content. We accidentally found through experiments that by adjusting the stirring speed, a relatively ideal drug loading concentration can be achieved without adding ethanol as a cosolvent. The stirring speed is one of the important core technologies of the present application, therefore we add the protection range of stirring speed in the claims.

[0355] Example 39: Effect of adjuvant ratio on dilution stability of nimodipine cyclodextrin inclusion complex

[0356] 1. Formulation

[0357] Table 23 Formulation design

[0358]

[0359] 2. Preparation process

[0360] Take 15 g, 17.5 g, 20 g, 22.5 g, 25 g, 30 g, 35 g of sulfobutyl ether-β-cyclodextrin respectively, add an appropriate amount of water for injection, stir to dissolve at 75°C to obtain a sulfobutyl ether-β-cyclodextrin aqueous solution; take 50 mg of nimodipine, under the condition of stirring speed of 1.8 m / s, add nimodipine to the sulfobutyl ether-β-cyclodextrin aqueous solution, stir for 1.5 h, add water for injection to make up to 100 mL, the obtained solution is sterile filtered through a 0.22 μm microporous filter, filled, freeze-dried, and packaged.

[0361] The prepared nimodipine for injection is diluted with 0.9% sodium chloride injection to a drug concentration of 0.2 mg / mL. At 0, 12, 24, 36, 48, and 60 h, observe and take samples, filter through a 0.45 μm polyether sulfone filter, and then determine the nimodipine content according to the above content determination method.

[0362] 3. Experimental results

[0363] The dilution stability results are shown in Table 24. When the auxiliary drug ratio is ≥350, the dilution stability time of nimodipine for injection can be up to 24 h or more; when the auxiliary drug ratio is ≥400, the dilution stability time of nimodipine for injection can be up to 36 h or more; when the auxiliary drug ratio is ≥450, the dilution stability time of nimodipine for injection can be up to 48 h or more; when the auxiliary drug ratio is ≥500, the dilution stability time of nimodipine for injection can be up to 60 h or more. The results show that the auxiliary drug ratio significantly affects the dilution stability of nimodipine cyclodextrin inclusion complex.

[0364] Table 24 Dilution stability of nimodipine cyclodextrin inclusion complex at different auxiliary drug ratios

[0365] Ratio of adjuvant Dilution stability 300 Content significantly reduced in 12h, stability time < 12h 350 Content almost unchanged in 24h, stability time > 24h 400 Content almost unchanged in 36h, 36h < stability time < 48h 450 Content almost unchanged in 48h, stability time > 48h 500 Content almost unchanged in 60h, stability time > 60h 600 Content almost unchanged in 60h, stability time > 60h 700 Content almost unchanged in 60h, stability time > 60h

[0366] 4. Discussion

[0367] The research results show that the dilution stability time of nimodipine cyclodextrin inclusion complex increases with the increase of the auxiliary drug ratio. According to the recommended dosage regimen in the instructions, the maximum specification of nimodipine injection (250 mL: 50 mg) has a maximum continuous administration time of 51 h. The research found that when the auxiliary drug ratio of nimodipine cyclodextrin inclusion complex is ≥500, the dilution stability time can be up to 60 h or more, meeting the clinical use requirements. Based on the above reasons, we increased the range of the mass ratio of sulfobutyl ether-β-cyclodextrin and nimodipine in the claims.

[0368] Example 40: Preparation of nimodipine for injection

[0369] 1. Prescription

[0370]

[0371] 2. Preparation process

[0372] Take sulfobutyl ether-β-cyclodextrin 3g, add the appropriate amount of water for injection, 70℃ stirring to dissolve, get sulfobutyl ether-β-cyclodextrin aqueous solution; take nimodipine 3mg, dissolved in 0.07ml ethanol, under the condition of stirring speed is 3.0m / s nimodipine ethanol solution is added to sulfobutyl ether-β-cyclodextrin aqueous solution, stirring 10min, add water for injection to 30ml, the resulting solution was sterile filtered by 0.22μm microporous filter, filling, freeze drying, packaging, namely.

[0373] Example 41: the preparation of nimodipine for injection

[0374] 1. Prescription

[0375]

[0376] 2. Preparation process

[0377] Take sulfobutyl ether-β-cyclodextrin 3g, add the appropriate amount of water for injection, 60℃ stirring to dissolve, get sulfobutyl ether-β-cyclodextrin aqueous solution; take nimodipine 6mg, under the condition of stirring speed is 2.0m / s nimodipine ethanol solution is added to sulfobutyl ether-β-cyclodextrin aqueous solution, stirring 30min, add water for injection to 30ml, the resulting solution was sterile filtered by 0.22μm microporous filter, filling, freeze drying, packaging, namely.

[0378] Example 42: the preparation of nimodipine for injection

[0379] 1. Prescription

[0380]

[0381] 2. Preparation process

[0382] Take sulfobutyl ether-β-cyclodextrin 3g, add the appropriate amount of water for injection, 20℃ stirring to dissolve, get sulfobutyl ether-β-cyclodextrin aqueous solution; take nimodipine 6mg, under the condition of stirring speed is 10.0m / s nimodipine is added to sulfobutyl ether-β-cyclodextrin aqueous solution, stirring 4h, add water for injection to 30ml, the resulting solution was sterile filtered by 0.22μm microporous filter, filling, freeze drying, packaging, namely.

[0383] Example 43: the preparation of nimodipine for injection

[0384] 1. Prescription

[0385]

[0386] 2. Preparation process

[0387] Take sulfobutyl ether-β-cyclodextrin 4.5 g, sodium citrate 60 mg and citric acid 9 mg, add an appropriate amount of water for injection, stir to dissolve at 90°C, get sulfobutyl ether-β-cyclodextrin aqueous solution; take nimodipine 11.25 mg, under the condition of stirring speed 0.5 m / s, nimodipine is added to the sulfobutyl ether-β-cyclodextrin aqueous solution, stirring for 3 h, add water for injection to 30 mL,

[0388] Example 44: Preparation of nimodipine for injection

[0389] 1. Prescription

[0390]

[0391] 2. Preparation process

[0392] Take sulfobutyl ether-β-cyclodextrin 6 g, add an appropriate amount of water for injection, stir to dissolve at 70°C, get sulfobutyl ether-β-cyclodextrin aqueous solution; take nimodipine 12 mg, under the condition of stirring speed 2.0 m / s, nimodipine is added to the sulfobutyl ether-β-cyclodextrin aqueous solution, stirring for 3 h, add water for injection to 30 mL, the resulting solution is sterile filtered through 0.22 μm microporous filter, fill, freeze-drying, nitrogen filling, packaging, ready.

[0393] Example 45: Preparation of nimodipine for injection

[0394] 1. Prescription

[0395]

[0396] 2. Preparation process

[0397] Take sulfobutyl ether-β-cyclodextrin 6 g, add an appropriate amount of water for injection, stir to dissolve at 85°C, get sulfobutyl ether-β-cyclodextrin aqueous solution; take nimodipine 12 mg, under the condition of stirring speed 0.8 m / s, nimodipine is added to the sulfobutyl ether-β-cyclodextrin aqueous solution, stirring for 2 h, add water for injection to 30 mL, the resulting solution is sterile filtered through 0.22 μm microporous filter, fill, freeze-drying, packaging, ready.

[0398] Example 46: Preparation of nimodipine for injection

[0399] 1. Prescription

[0400]

[0401]

[0402] 2. Preparation process

[0403] Take sulfobutyl ether-β-cyclodextrin 6.3g, add an appropriate amount of water for injection, stirring to dissolve at 40℃, sulfobutyl ether-β-cyclodextrin aqueous solution; take nimodipine 14mg, under the condition of stirring speed 6.0m / s nimodipine is added to the sulfobutyl ether-β-cyclodextrin aqueous solution, stirring 3h, add water for injection to 30mL, the resulting solution is sterile filtered through 0.22μm microporous filter, filling, freeze-drying, packaging, ready.

[0404] Example 47: Preparation of nimodipine for injection

[0405] 1. Prescription

[0406]

[0407] 2. Preparation process

[0408] Take sulfobutyl ether-β-cyclodextrin 6.6g, add an appropriate amount of water for injection, stirring to dissolve at 60℃, sulfobutyl ether-β-cyclodextrin aqueous solution; take nimodipine 14.67mg, dissolved in 0.3mL ethanol, under the condition of stirring speed 2.0m / s nimodipine ethanol solution is added to the sulfobutyl ether-β-cyclodextrin aqueous solution, stirring 2h, add water for injection to 30mL, the resulting solution is sterile filtered through 0.22μm microporous filter, filling, freeze-drying, nitrogen filling, packaging, ready. The whole process of dissolution and filtration is carried out under clean and dry N2.

[0409] Example 48: Preparation of nimodipine for injection

[0410] 1. Prescription

[0411]

[0412] 2. Preparation process

[0413] Take sulfobutyl ether-β-cyclodextrin 7.5g, add an appropriate amount of water for injection, stirring to dissolve at 75℃, sulfobutyl ether-β-cyclodextrin aqueous solution; take nimodipine 15mg, under the condition of stirring speed 1.8m / s nimodipine is added to the sulfobutyl ether-β-cyclodextrin aqueous solution, stirring 1.5h, add water for injection to 30mL, the resulting solution is sterile filtered through 0.22μm microporous filter, filling, freeze-drying, packaging, ready.

[0414] Example 49: Preparation of nimodipine for injection

[0415] 1. Prescription

[0416]

[0417] 2. Preparation process

[0418] Take sulfobutyl ether-β-cyclodextrin 7.5 g, add an appropriate amount of water for injection, 80 ℃ stirring to dissolve, get sulfobutyl ether-β-cyclodextrin aqueous solution; take nimodipine 15 mg, under the condition of stirring speed 1.7 m / s nimodipine is added to the sulfobutyl ether-β-cyclodextrin aqueous solution, stirring 2 h, add water for injection to 30 mL, the resulting solution is sterile filtered through 0.22 μm microporous filter, filling, freeze-drying, packaging, get.

[0419] Example 50: Preparation of nimodipine for injection

[0420] 1. Prescription

[0421]

[0422] 2. Preparation process

[0423] Take sulfobutyl ether-β-cyclodextrin 7.5 g, add an appropriate amount of water for injection, 90 ℃ stirring to dissolve, get sulfobutyl ether-β-cyclodextrin aqueous solution; take nimodipine 15 mg, under the condition of stirring speed 1.5 m / s nimodipine is added to the sulfobutyl ether-β-cyclodextrin aqueous solution, stirring 1 h, add water for injection to 30 mL, the resulting solution is sterile filtered through 0.22 μm microporous filter, filling, freeze-drying, packaging, get.

[0424] Example 51: Preparation of nimodipine for injection

[0425] 1. Prescription

[0426]

[0427] 2. Preparation process

[0428] Take sulfobutyl ether-β-cyclodextrin 7.5 g, add an appropriate amount of water for injection, 75 ℃ stirring to dissolve, get sulfobutyl ether-β-cyclodextrin aqueous solution; take nimodipine 15 mg, under the condition of stirring speed 1.9 m / s nimodipine is added to the sulfobutyl ether-β-cyclodextrin aqueous solution, stirring 1.5 h, add water for injection to 30 mL, the resulting solution is sterile filtered through 0.22 μm microporous filter, filling, freeze-drying, packaging, get.

[0429] Example 52: Preparation of nimodipine for injection

[0430] 1. Prescription

[0431]

[0432] 2. Preparation process

[0433] Take sulfobutyl ether-β-cyclodextrin 7.5 g, add the appropriate amount of water for injection, stirring to dissolve at 65 ℃, sulfobutyl ether-β-cyclodextrin aqueous solution; nimodipine 15 mg, under the condition of stirring speed of 2.0 m / s nimodipine added to sulfobutyl ether-β-cyclodextrin aqueous solution solution, stirring 2 h, add water for injection to 30 mL, the resulting solution was sterile filtered through 0.22 μm microporous membrane, filling, freeze-drying, packaging, ready.

[0434] Example 53: the preparation of nimodipine for injection

[0435] 1. prescription

[0436]

[0437] 2. preparation process

[0438] Take sulfobutyl ether-β-cyclodextrin 7.5 g, add the appropriate amount of water for injection, stirring to dissolve at 75 ℃, sulfobutyl ether-β-cyclodextrin aqueous solution; nimodipine 15 mg, under the condition of stirring speed of 2.5 m / s nimodipine added to sulfobutyl ether-β-cyclodextrin aqueous solution solution, stirring 3 h, add water for injection to 30 mL, the resulting solution was sterile filtered through 0.22 μm microporous membrane, filling, freeze-drying, packaging, ready.

[0439] Example 54: the preparation of nimodipine for injection

[0440] 1. prescription

[0441]

[0442] 2. preparation process

[0443] Take sulfobutyl ether-β-cyclodextrin 7.5 g, add the appropriate amount of water for injection, stirring to dissolve at 60 ℃, sulfobutyl ether-β-cyclodextrin aqueous solution; nimodipine 15 mg, under the condition of stirring speed of 3.0 m / s nimodipine added to sulfobutyl ether-β-cyclodextrin aqueous solution solution, stirring 3 h, add water for injection to 30 mL, the resulting solution was sterile filtered through 0.22 μm microporous membrane, filling, freeze-drying, packaging, ready.

[0444] Example 55: the preparation of nimodipine for injection

[0445] 1. prescription

[0446]

[0447] 2. preparation process

[0448] Take sulfobutyl ether-β-cyclodextrin 7.5 g, add an appropriate amount of water for injection, 70°C stirring to dissolve, get sulfobutyl ether-β-cyclodextrin aqueous solution; take nimodipine 15 mg, dissolved in 0.45 mL of ethanol, under the condition of stirring speed is 2.0 m / s nimodipine is added to sulfobutyl ether-β-cyclodextrin aqueous solution, stirring 1 h, add water for injection to 30 mL, the resulting solution was sterile filtered by 0.22 μm microporous filter, filling, freeze-drying, packaging, get.

[0449] Example 56: preparation of nimodipine for injection

[0450] 1. prescription

[0451]

[0452] 2. preparation process

[0453] Take sulfobutyl ether-β-cyclodextrin 9 g, add an appropriate amount of water for injection, 90°C stirring to dissolve, get sulfobutyl ether-β-cyclodextrin aqueous solution; take nimodipine 18 mg, dissolved in 0.75 mL of ethanol, under the condition of stirring speed is 1.0 m / s nimodipine ethanol solution is added to sulfobutyl ether-β-cyclodextrin aqueous solution, stirring 3 h, add water for injection to 30 mL, the resulting solution was sterile filtered by 0.22 μm microporous filter, filling, freeze-drying, packaging, get.

[0454] Example 57: preparation of nimodipine for injection

[0455] 1. prescription

[0456]

[0457] 2. preparation process

[0458] Take sulfobutyl ether-β-cyclodextrin 9 g, add an appropriate amount of water for injection, 75°C stirring to dissolve, get sulfobutyl ether-β-cyclodextrin aqueous solution; take nimodipine 18 mg, under the condition of stirring speed is 3.0 m / s nimodipine is added to sulfobutyl ether-β-cyclodextrin aqueous solution, stirring 2 h, add water for injection to 30 mL, the resulting solution was sterile filtered by 0.22 μm microporous filter, filling, freeze-drying, nitrogen, packaging, get. The whole process of dissolution and filtration into clean dry N2.

[0459] Example 58: preparation of nimodipine for injection

[0460] 1. prescription

[0461]

[0462] 2. preparation process

[0463] Take sulfobutyl ether-β-cyclodextrin 10.5 g, add an appropriate amount of water for injection, stirring to dissolve at 40°C, sulfobutyl ether-β-cyclodextrin aqueous solution; take nimodipine 19.10 mg, under the condition of stirring speed 6.0 m / s nimodipine is added to the sulfobutyl ether-β-cyclodextrin aqueous solution, stirring 5 h, add water for injection to 30 mL, the resulting solution is sterile filtered through a 0.22 μm microporous filter, fill, freeze-drying, packaging, ready.

[0464] Example 59: Preparation of nimodipine for injection

[0465] 1. Prescription

[0466]

[0467] 2. Preparation process

[0468] Take sulfobutyl ether-β-cyclodextrin 11.4 g, potassium dihydrogen phosphate 204 mg and sodium hydroxide 3.5 mg, add an appropriate amount of water for injection, stirring to dissolve at 90°C, sulfobutyl ether-β-cyclodextrin aqueous solution; take nimodipine 20.72 mg, under the condition of stirring speed 1.0 m / s nimodipine is added to the sulfobutyl ether-β-cyclodextrin aqueous solution, stirring 2 h, add water for injection to 30 mL, the resulting solution is sterile filtered through a 0.22 μm microporous filter, fill, freeze-drying, packaging, ready.

[0469] Example 60: Preparation of nimodipine for injection

[0470] 1. Prescription

[0471]

[0472] 2. Preparation process

[0473] Take sulfobutyl ether-β-cyclodextrin 12 g, add an appropriate amount of water for injection, stirring to dissolve at 95°C, sulfobutyl ether-β-cyclodextrin aqueous solution; take nimodipine 30 mg, under the condition of stirring speed 0.8 m / s nimodipine is added to the sulfobutyl ether-β-cyclodextrin aqueous solution, stirring 4 h, add water for injection to 30 mL, the resulting solution is sterile filtered through a 0.22 μm microporous filter, fill, freeze-drying, packaging, ready.

[0474] Example 61: Preparation of nimodipine for injection

[0475] 1. Prescription

[0476]

[0477] 2. Preparation process

[0478] Take sulfobutyl ether-β-cyclodextrin 12 g, add the appropriate amount of water for injection, stirring to dissolve at 80°C, sulfobutyl ether-β-cyclodextrin aqueous solution; take nimodipine 30 mg, under the condition of stirring speed of 2.0 m / s nimodipine is added to the sulfobutyl ether-β-cyclodextrin aqueous solution, stirring 3 h, add water for injection to 30 mL, the resulting solution is sterile filtered through 0.22 μm microporous filter, fill, freeze-drying, nitrogen, packaging, ready. The whole process of dissolution and filtration into clean dry N2.

[0479] Example 62: Preparation of nimodipine for injection

[0480] 1. Prescription

[0481]

[0482] 2. Preparation process

[0483] Take sulfobutyl ether-β-cyclodextrin 13.5 g, add the appropriate amount of water for injection, stirring to dissolve at 75°C, sulfobutyl ether-β-cyclodextrin aqueous solution; take nimodipine 22.5 mg, under the condition of stirring speed of 3.0 m / s nimodipine is added to the sulfobutyl ether-β-cyclodextrin aqueous solution, stirring 2 h, add water for injection to 30 mL, the resulting solution is sterile filtered through 0.22 μm microporous filter, fill, freeze-drying, packaging, ready.

[0484] Example 63: Preparation of nimodipine for injection

[0485] 1. Prescription

[0486]

[0487] 2. Preparation process

[0488] Take sulfobutyl ether-β-cyclodextrin 15 g, add the appropriate amount of water for injection, stirring to dissolve at 100°C, sulfobutyl ether-β-cyclodextrin aqueous solution; take nimodipine 42 mg, dissolved in 1.05 mL of ethanol, under the condition of stirring speed of 1.7 m / s nimodipine ethanol solution is added to the sulfobutyl ether-β-cyclodextrin aqueous solution, stirring 1.5 h, add water for injection to 30 mL, the resulting solution is sterile filtered through 0.22 μm microporous filter, fill, freeze-drying, packaging, ready.

[0489] Example 64: Preparation of nimodipine for injection

[0490] 1. Prescription

[0491]

[0492] 2. Preparation process

[0493] Sulfobutyl ether-β-cyclodextrin 15 g, add an appropriate amount of water for injection, 95 ℃ stirring to dissolve, get sulfobutyl ether-β-cyclodextrin aqueous solution; take nimodipine 21.43 mg, under the condition of stirring speed 2.0 m / s, nimodipine is added to sulfobutyl ether-β-cyclodextrin aqueous solution, stirring for 2 h, add water for injection to 30 mL, the resulting solution is sterile filtered by 0.22 μm microporous filter membrane, filling, freeze-drying, nitrogen filling, packaging, get. The whole dissolving and filtering process is carried out under clean and dry N2.

[0494] Example 65: Ethanol residue determination of nimodipine for injection

[0495] 1. Experimental method

[0496] Select nimodipine for injection prepared in examples 40, 41, 47, 55, 56, 63, and detect ethanol residue. The ethanol content in nimodipine for injection is determined according to residual solvent determination method (Chinese Pharmacopoeia 2020 edition four general rules 0861 second method).

[0497] 2. Experimental results

[0498] Table 25 ethanol residue results of nimodipine for injection

[0499] Sample name Ethanol residue (%) Nimodipine for injection (Example 40) 0.03 Nimodipine for injection (Example 41) 0.03 Nimodipine for injection (Example 47) 0.04 Nimodipine for injection (Example 55) 0.06 Nimodipine for injection (Example 56) 0.09 Nimodipine for injection (Example 63) 0.15

[0500] The results show that by controlling the temperature and time, the ethanol residue of nimodipine for injection of the application is lower.

[0501] Example 66: Dilution stability study of nimodipine for injection

[0502] 1. Experimental method

[0503] Take nimodipine for injection prepared in example 48, add water for injection for reconstitution, and nimodipine injection, respectively, dilute to the clinical drug concentration of 0.04 mg / mL with 0.9% sodium chloride injection and 5% glucose. At 0, 3, 6, 9, 12, 24 h, observe and take samples, filter with 0.45 μm polyether sulfone filter membrane, and determine the content of nimodipine according to the above content determination method.

[0504] 2. Experimental results

[0505] The dilution stability results of nimodipine for injection and nimodipine injection diluted with 0.9% sodium chloride injection are shown in tables 26 and 27, and the solution appearance is shown in Figure 4 .

[0506] Table 26 content change of nimodipine for injection and nimodipine injection diluted with 0.9% sodium chloride injection

[0507]

[0508]

[0509] Note: "-" indicates that the drug is not precipitated, "+" indicates that the drug is precipitated.

[0510] Table 27 Content change of nimodipine for injection and nimetop injection diluted with 5% glucose injection

[0511]

[0512] Note: "-" indicates that the drug is not precipitated, "+" indicates that the drug is precipitated.

[0513] After nimetop injection is diluted with 0.9% sodium chloride injection or 5% glucose injection, the drug is precipitated in a large amount within 3h, and the content of the drug is significantly reduced within 24h, so the dilution stability is very poor; after the nimodipine for injection of the application is diluted with 0.9% sodium chloride injection or 5% glucose injection, no obvious change in content occurs within 24h, and the appearance is clear, so the dilution stability is good. The above results show that compared with nimetop injection, the dilution stability of the nimodipine for injection of the application is significantly improved, and has obvious advantages.

[0514] Example 67: Influencing factors and accelerated stability investigation of nimodipine for injection

[0515] 1. Experimental method

[0516] The nimodipine for injection prepared in Example 48 was selected, and the high temperature, high humidity, light and accelerated test stability of the nimodipine for injection was investigated according to the “9001 Raw Materials and Preparation Stability Test Guiding Principles” in the 2020 edition of “People's Republic of China Pharmacopoeia”, and the changes of drug content and related substances were detected.

[0517] 2. Experimental results

[0518] Table 28 Stability investigation results of nimodipine for injection

[0519]

[0520] 3. Results analysis

[0521] Under the conditions of high temperature, high humidity and acceleration test, the content of nimodipine for injection and the related substances almost do not change. But under the condition of light intensity for 10 days, the content of nimodipine for injection is only 96.59%, and the related substances increase by 0.11%. It may be that nimodipine is less stable under light conditions and is easy to decompose, even after being wrapped by cyclodextrin, it is still degraded under light conditions, indicating that nimodipine for injection should be stored in the dark. The above results show that the nimodipine for injection of the application has good stability, and the quality meets the relevant provisions.

[0522] Example 68: Vessel irritability study of nimodipine for injection

[0523] 1. Experimental method

[0524] Six rabbits (body weight 1.8-2.0 kg) were randomly divided into two groups. The nimodipine for injection prepared in Example 48 was dissolved in water for injection, and the commercial nimodipine injection was diluted to a clinical administration concentration of 0.04 mg / mL with 0.9% sodium chloride injection. The drug solution was injected into the right auricular vein of the rabbits at a dose of 0.4 mg / kg and a push injection speed of 1 mL / min. At the same time, the left ear was given an equal volume of 0.9% sodium chloride injection as a control. This was done once a day for three consecutive days. During the administration period, the animal behavior and changes at the injection site were observed macroscopically. Twenty-four to forty-eight hours after the last administration, the animals were euthanized, and the auricular vein tissue about 0.5-3.0 cm from the injection site was taken and fixed in 10% paraformaldehyde solution. Ethanol gradient dehydration was used, and the tissue was embedded in paraffin. Hematoxylin-eosin staining was performed, and the pathological changes were evaluated.

[0525] 2. Experimental results

[0526] The results of the rabbit ear pathological sections of the vessel irritability are shown in Table 1. Figure 5

[0527] The nimodipine injection showed obvious collagen swelling, red blood cell leakage, and punctate hemorrhage, with endothelial cell swelling and a large number of inflammatory cell infiltration, indicating that the nimodipine injection group had a strong inflammatory response after administration. The rabbits struggled violently and had a strong pain sensation during the administration process, which may be due to the strong vessel irritability and pain sensation caused by high-concentration ethanol entering the blood vessels.

[0528] The auricular skin surface epithelial structure of the nimodipine for injection of the present application and 0.9% sodium chloride injection was complete, and there was no special lesion in the sebaceous gland and sweat gland tissue. The cartilage morphology in the central part was normal, and no obvious lesion damage was observed. During the administration process, the rabbits had no obvious discomfort. Compared with the 0.9% sodium chloride injection group, the nimodipine for injection had only a small amount of inflammatory cell infiltration and no other abnormal conditions.

[0529] The above results show that the vessel irritability of the nimodipine for injection of the present application is significantly lower than that of the nimodipine injection, and the safety is improved.

[0530] Example 69: Hemolytic study of nimodipine for injection

[0531] 1. Experimental method

[0532] ​Nimodipine for injection (NIMO-CD) prepared in Example 48 was reconstituted with water for injection and commercially available Nimotop injection was diluted with 0.9% sodium chloride injection to a clinically administered concentration of 0.04 mg / mL. Eight clean 10 mL glass tubes were taken and numbered: tubes 1 to 5 contained different concentrations of NIMO-CD, tube 6 was the negative control, tube 7 was the positive control, and tube 8 contained Nimotop injection. As shown in the table below, 2% red blood cell suspension, 0.9% sodium chloride injection, distilled water, and the drug solution were added sequentially. After mixing, the tubes were immediately incubated in a water bath at 37±0.5℃. The hemolysis status of each tube was observed and recorded. Initially, records were taken every 15 minutes, then every hour after 1 hour, for a total of 3 hours.

[0533] Table 29 Hemolysis Experimental Design

[0534]

[0535] 2. Experimental Results

[0536] Hemolysis such as Figure 6 As shown, NIMO-CD (types 1-5) red blood cells all deposited at the bottom of the test tube, and the supernatant was colorless and clear. After shaking, all cells dispersed evenly, indicating that NIMO-CD does not cause hemolysis or red blood cell aggregation. These results demonstrate that the nimodipine for injection of this invention is non-hemolytic and suitable for injection.

[0537] Example 70: Pharmacodynamic Study of Nimodipine for Injection

[0538] 1. Experimental Methods

[0539] 1.1 Grouping and administration of experimental animals

[0540] Thirty-two male SD rats were randomly divided into four groups: sham-operated group (Sham group), model control group (Model group), Nimotop injection group (Nimotop group), and nimodipine injection group (NIMO-CD group, Example 48). Nimodipine injection prepared in Example 48 was reconstituted with water for injection and then diluted to 0.2 mg / mL with 0.9% sodium chloride injection. The Nimotop and NIMO-CD groups were administered the drug via tail vein injection at a dose of 1 mg / kg, once daily from the day of modeling until 72 hours post-surgery, and immediately after surgery.

[0541] 1.2 Establishment of a cerebral hemorrhage model

[0542] The rats in the Model group, Nimotop group and NIMO-CD group were anesthetized with 1-4% isoflurane, and a brain stereotaxic instrument was used to inject 2 mg / mL collagenase solution 2 pL at a rate of 1 pL / 5 min into the left striatum area (centered on the bregma, 2.0 mm posterior, 2.0 mm right, and 4.0 mm deep) for 2 min, and then slowly removed after 3 min of needle residence to establish a cerebral hemorrhage model. After the operation, the wound was sutured and disinfected with iodophor. The hind limb was injected with 40 mg / kg gentamicin to prevent infection, and 10 mg / kg analgin was injected intramuscularly twice a day to relieve pain. The animals in the Sham group were subjected to the same operation as above, and the same volume of PBS solution was used instead of collagenase injection.

[0543] 1.3 Detection index

[0544] 1.3.1 Behavioral evaluation

[0545] Before the operation and at 24 h, 48 h and 72 h after the establishment of the cerebral hemorrhage model, the animal behavior was observed, and the modified neurological severity score (mNSS) was used to evaluate the motor, sensory, balance and reflex abilities of the experimental animals. The corner test (CT) was used to measure the sensory motor dysfunction of the experimental animals. The rats were placed in a 30° angle formed by two wooden boards, and the number of times of turning to the right was observed and recorded. The test was performed for 10 times, and the CT score was calculated according to the following formula.

[0546]

[0547] 1.3.2 Cerebral hemorrhage injury area

[0548] At the end of the experiment, all experimental animals were anesthetized and euthanized by intraperitoneal injection of 25-50 mg / kg Xuelai. The brain was taken out, fixed in 4% paraformaldehyde solution for 24 h, and then placed in a brain mold. The brain was cut into 6 pieces from front to back in a coronal manner with the needle insertion position as the center. The 6 brain slices were arranged in order and imaged with a digital camera. The area ratio of each slice was calculated by image analysis software ImageJ (version: 1.4.3.67), and the total hematoma area difference between groups was compared. If an animal died during the experiment, the brain was immediately taken out and processed according to the above procedure.

[0549] 1.3.3 Histopathological evaluation

[0550] After the rat brain tissue slices were fixed in 4% paraformaldehyde solution, paraffin-embedded and HE-stained, the slice pathological changes were observed under an optical microscope. The brain edema, inflammatory infiltration and neuronal damage of the slice were evaluated according to the histopathological scoring standard, and the scoring standard is shown in Table 30.

[0551] Table 30 Histopathology scoring criteria

[0552]

[0553] 2. Experimental results

[0554] 2.1 Behavioral evaluation

[0555] The mNSS scores are shown in Table 31 and Figure 7 A, the international standard definition of mNSS score: normal SD rats are 0 points, 1-6 points for mild injury, 7-12 points for moderate injury, 13-18 points for severe injury. The mNSS score of the Model group was the highest on the first day, with the most severe neurological impairment, and the scores gradually decreased on the second and third days, with gradual recovery of neurological function, but were all greater than 7 points, showing moderate injury. The neurological function scores of the NIMO-CD group and the Nimotop group also decreased over time, and the scores were greater than 7 points on the two days after treatment, showing moderate injury, and were less than 7 points on the third day, showing mild injury. Compared with the Model group, the mNSS scores of the NIMO-CD group and the Nimotop group were significantly reduced.

[0556] Table 31 mNSS score table (Mean ± SD, n = 8)

[0557]

[0558] Note: compared with the Model group, *P<0.05, **P<0.01, ***P<0.001.

[0559] The CT score results are shown in Table 32 and Figure 7 B, the CT score decreased over time, and the experimental animals in each group gradually improved their hemiplegia. The final CT scores of the Sham group and the Model group were 48.75% and 65.00%, respectively, and the final CT scores of the NIMO-CD group and the Nimotop group were 58.75% and 60.00%, respectively. Compared with the Model group, the CT scores of the NIMO-CD group and the Nimotop group were reduced.

[0560] Table 32 CT score table (Mean ± SD, n = 8)

[0561]

[0562] Note: compared with the Model group, *P<0.05, **P<0.01, ***P<0.001.

[0563] Behavioral evaluation results showed that nimodipine injection significantly improved neurobehavioral dysfunction caused by brain injury in a rat model of cerebral hemorrhage, and there was no significant difference compared with the nimodipine injection group (P>0.05).

[0564] 2.2 Area of ​​brain hemorrhage injury

[0565] Brain tissue was collected from the experimental animals, and the differences in total hematoma area among the groups were compared. The results are shown in Table 33 and... Figure 7 As shown in Figure C, no hematoma was observed in the Sham group, the total hematoma area in the Model group was 4.45%, and the total hematoma area in both the NIMO-CD and Nimotop groups was 0.98%. Compared with the Model group, the total hematoma area in the NIMO-CD and Nimotop groups was significantly reduced. The results of the brain hemorrhage injury area measurement experiment showed that nimodipine injection significantly reduced hematoma caused by brain hemorrhage in rats and significantly improved brain hemorrhage injury, with no significant difference compared to nimodipine injection (P>0.05).

[0566] Table 33. Injury area of ​​cerebral hemorrhage (Mean±SD, n=8)

[0567]

[0568] Note: Compared with the Model group, *P<0.05, **P<0.01, ***P<0.001.

[0569] 2.3 Histopathological evaluation

[0570] Microscopic observation and analysis of brain tissue sections, pathological sections such as... Figure 7 As shown in E, the degree of cerebral edema, inflammation, and neuronal damage were comprehensively scored based on pathological sections. The scoring results are shown in Table 34 and... Figure 7 As shown in Figure D, the histopathological evaluation scores for the Sham group and the Model group were 1.38 and 5.00, respectively. The histopathological evaluation scores for the Nimotop group and the NIMO-CD group were 2.63 and 2.25, respectively. Compared with the Model group, the total histopathological evaluation score of the experimental animals in the NIMO-CD group was lower. Histopathological results indicate that nimodipine injection significantly improved the degree of cerebral edema, inflammation, and neuronal damage, and there was no significant difference compared with nimodipine injection (P>0.05).

[0571] Table 34 Histopathological Scoring Scale (Mean±SD, n=8)

[0572]

[0573] Note: Compared with the Model group, *P<0.05, **P<0.01, ***P<0.001.

[0574] In summary, the injection of nimodipine has a significant therapeutic effect on the prognosis of cerebral hemorrhage, greatly improving the degree of brain hematoma, brain edema, inflammation and neuronal damage caused by cerebral hemorrhage, and the treatment effect is basically the same as that of nimodipine injection.

[0575] Example 71: Pharmacokinetic study of nimodipine for injection

[0576] 1. Experimental method

[0577] 1.1 Test animal grouping and administration

[0578] 16 male SD rats were randomly divided into two groups, 8 rats in each group, and fasted for 12 h before the experiment. The nimodipine for injection (NIMO-CD) prepared in Example 48 was added to the injection water and then diluted to 0.2 mg / mL with 0.9% sodium chloride injection. 1.6 mg / kg of nimodipine injection (Nimotop) and NIMO-CD were administered through the tail vein, respectively, and 0.5 mL of blood was taken from the orbit at 5, 15, 30, 45, 60, 120, 240, 360 and 480 min after administration. 4000 rpm centrifugation for 5 min, take 300 μL of supernatant, -80℃ refrigerator frozen preservation.

[0579] 1.2 Blood plasma sample processing method

[0580] After thawing the frozen plasma samples at room temperature, centrifuge at 4000 rpm for 5 min, take 200 μL of plasma and place it in a 2.0 mL EP tube, add 40 μL of 5 μg / mL felodipine internal standard solution, vortex for 5 min. Add 100 μL of 1 mol / L NaOH, quantitatively add 1.5 mL of anhydrous ether: n-hexane (1:1) extraction solution, vortex for 5 min, ultrasonic for 5 min, centrifuge at 10000 rpm for 10 min, take all the supernatant in a 1.5 mL EP tube, blow dry at 45℃, the residue is dissolved with 100 μL of mobile phase, vortex mix for 5 min, centrifuge at 10000 rpm for 10 min, HPLC injection for determination.

[0581] 1.3 Chromatographic conditions

[0582] Chromatographic column: Zorbax SB C 18 column (250 mm x 4.6 mm, 5 μm)

[0583] Mobile phase: acetonitrile: water (60:40, v / v)

[0584] Flow rate: 1.0 mL / min

[0585] Detection wavelength: 358 nm

[0586] Column temperature: 30 °C

[0587] Injection volume: 20 μL

[0588] 2. Experimental results

[0589] The blood concentration-time curves of SD rats after tail vein injection of Nimotop injection and NIMO-CD are shown in Figure 8. As shown in Figure 8, the pharmacokinetic elimination curves of NIMO-CD and Nimotop injection in SD rats after intravenous administration are almost the same. NIMO enters the body and is rapidly eliminated within a short time after administration of NIMO-CD and Nimotop injection. HPLC cannot detect the drug in the plasma 240 min after administration. Figure 8

[0590] The main pharmacokinetic parameters of NIMO-CD and Nimotop injection are shown in Table 35. The results show that the pharmacokinetic parameters of NIMO-CD and Nimotop injection are basically the same, including the area under the blood concentration-time curve (AUC), mean residence time (MRT), half-life (t 1 / 2 ), clearance (CL) and maximum blood drug concentration (C max ), and the differences between groups have no statistical significance (P>0.05). The above results show that the pharmacokinetic properties of Nimodipine for injection and Nimotop injection are basically the same after tail vein injection in SD rats.

[0591] Table 35 Main pharmacokinetic parameters of NIMO-CD and Nimotop injection (Mean ± SD, n=8)

[0592]

[0593] The above has specifically described the preferred embodiments of the present application, but the present application is not limited to the described embodiments, and those skilled in the art can make various equivalent modifications or replacements without departing from the spirit of the present application. These equivalent modifications or replacements are all included in the scope defined by the claims of the present application.​

Claims

1. A safe, stable, injectable nimodipine characterized in that, Sulfobutyl ether-β-cyclodextrin and nimodipine, the mass ratio of sulfobutyl ether-β-cyclodextrin and nimodipine is 350:1-700:1; The preparation method comprises the following steps: (a) weighing the prescribed amount of sulfobutyl ether-β-cyclodextrin, adding an appropriate amount of water for injection, stirring to dissolve at a certain temperature to obtain a sulfobutyl ether-β-cyclodextrin aqueous solution; the stirring temperature is 60-90°C; (b) under the conditions of a certain stirring temperature and speed, the nimodipine drug powder is added to the solution of step (a), stirred for a certain time, added with water for injection to the full amount, sterilized by 0.22 μm microporous filter membrane, filled, freeze-dried, packaged, and obtained; the stirring temperature is 60-90°C; the stirring time is 60-300 min; the stirring linear speed is 0.5-10.0 m / s.

2. The nimodipine for injection according to claim 1, wherein The nimodipine content of the nimodipine for injection before freeze-drying is 0.01-0.14% g / ml, and the sulfobutyl ether-β-cyclodextrin content is 10-50% g / ml.

3. The nimodipine for injection according to claim 2, wherein The mass ratio of sulfobutyl ether-β-cyclodextrin and nimodipine is 400:1-600:1, and the nimodipine content of the nimodipine for injection before freeze-drying is 0.02-0.10% g / ml, and the sulfobutyl ether-β-cyclodextrin content is 10-40% g / ml.

4. The nimodipine for injection according to claim 2, wherein The mass ratio of sulfobutyl ether-β-cyclodextrin and nimodipine is 450:1-550:1, and the nimodipine content of the nimodipine for injection before freeze-drying is 0.04-0.06% g / ml, and the sulfobutyl ether-β-cyclodextrin content is 20-30% g / ml.

5. A safe and stable nimodipine injection, characterized by, The nimodipine for injection is prepared from the following components: The mass ratio of sulfobutyl ether-β-cyclodextrin and nimodipine is 350:1-700:1; The preparation method comprises the following steps: (a) weighing the prescribed amount of sulfobutyl ether-β-cyclodextrin, adding an appropriate amount of water for injection, stirring to dissolve at a certain temperature to obtain a sulfobutyl ether-β-cyclodextrin aqueous solution; the stirring temperature is 60-90°C; (b) weighing the prescribed amount of nimodipine, adding the prescribed amount of ethanol to dissolve, and obtaining a nimodipine ethanol solution; (c) under the conditions of a certain stirring temperature and speed, the solution of step (b) or the nimodipine drug powder is added to the solution of step (a), stirred for a certain time, added with water for injection to the full amount, sterilized by 0.22 μm microporous filter membrane, filled, freeze-dried, packaged, and obtained; the stirring temperature is 60-90°C; the stirring time is 60-300 min; the stirring linear speed is 0.5-10.0 m / s.

6. The safe, stable injectable nimodipine of claim 5, wherein, The nimodipine for injection is prepared from the following components:

7. A process for the preparation of a safe, stable injection of nimodipine as claimed in claim 5 or 6, characterized in that, The preparation method comprises the following steps: (a) weighing the prescribed amount of sulfobutyl ether-β-cyclodextrin, adding an appropriate amount of water for injection, stirring to dissolve at a certain temperature to obtain a sulfobutyl ether-β-cyclodextrin aqueous solution; the stirring temperature is 60-90°C; (b) weighing the prescribed amount of nimodipine, adding the prescribed amount of ethanol to dissolve, and obtaining a nimodipine ethanol solution; (c) under certain stirring temperature and speed conditions, the solution of step (b) or nimodipine drug powder is added into the solution of step (a), stirred for a certain time, added with water for injection to the full amount, sterilized by 0.22 μm microporous filter, filled, freeze-dried, packaged, and the safe and stable nimodipine injection is obtained; the stirring temperature is 70-90 ℃; the stirring time is 60-300 min; and the stirring line speed is 0.5-10.0 m / s.

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